Cell production of pure iron oxide nanoparticles

Abstract

The present invention relates to the cellular production of pure iron oxide nanoparticles. Specifically, the present invention discloses a method for producing high-purity iron oxide nanoparticles using nanoparticle-producing cells, comprising: a) a pre-growth stage, including amplifying nanoparticle-producing cells in a pre-growth and / or fed-batch medium, and b) a growth stage, including amplifying nanoparticle-producing cells derived from the pre-growth stage in a growth and / or fed-batch medium, wherein each kilogram or liter of the pre-growth and / or growth and / or fed-batch medium contains: i) no more than 0.005 grams of yeast extract, and ii) no more than 0.001 grams of a CMR reagent selected from the group consisting of boric acid and nitrilotriacetic acid.

Description

Field of the Invention

[0001] The field of the invention is the bioproduction of nanoparticles containing low amounts of impurities. Background of the Invention

[0002] Bacteria that produce nanoparticles, such as magnetotactic bacteria, are known to accumulate impurities in their crystal structure. For example, when magnetotactic bacteria are grown in the presence of cobalt, they produce magnetosomes containing iron oxide and cobalt (S. Staniland et al, Nature nanotechnology, V. 3, P. 158 (2008)). For medical applications, it is desirable for the nanoparticles to contain low levels of toxic impurities, such as cobalt. Summary of the Invention

[0003] The present invention relates to a method for producing high-purity iron oxide nanoparticles using nanoparticle-producing cells, comprising:

[0004] a) a pre-growth phase, comprising amplifying nanoparticle-producing cells in a pre-growth and / or fed-batch culture medium (medium), preferably such that the nanoparticle-producing cells produce substantially no nanoparticles, and

[0005] b) a growth phase, comprising amplifying nanoparticle-producing cells derived from the pre-growth phase in a growth and / or fed-batch culture medium, preferably such that the nanoparticle-producing cells produce nanoparticles,

[0006] wherein each kilogram or liter of the pre-growth and / or growth and / or fed-batch culture medium contains:

[0007] i) no more than 5·10 5 、5·10 3 、50、5、0.5、0.005、0.0005、0.00005 or 5·10 -10 g of yeast extract, and / or

[0008] ii) no more than 10 5 、10 3 、10、1、0.1、0.001、10 -5 or 10 -10 g of CMR reagent, preferably selected from the group consisting of boric acid and nitrilotriacetic acid,

[0009] wherein, when a fed-batch culture medium is present, the fed-batch culture medium is preferably used as a medium to supplement the pre-growth and / or growth medium,

[0010] Among them, more nanoparticles are produced in the growth stage than in the pre-growth stage, preferably at least 0, 0.1, 0.5, 2, 5, 10, 10 3 、10 5 or 10 10 times, where the multiple is preferably Q2 / Q1, where Q1 and Q2 are preferably the nanoparticles produced in the pre-growth stage and the growth stage or sub-stage, respectively,

[0011] and / or

[0012] Among them, preferably the pre-growth and / or growth and / or fed-batch culture medium contains at least one other compound, which preferably participates in bacterial growth and / or magnetosome production or synthesis, and preferably the other compound is a carbon source, a nitrogen source, a calcium source, a vitamin source, an oxygen source, an iron source, a phosphate source, a phosphorus source, and / or a magnesium source. Description of the Drawings

[0013] Figure 1 : TGA-DSC analysis of whole magnetotactic bacteria and magnetosomes extracted from magnetotactic bacteria according to lysis condition 2. (a) For a sample containing 3 mg of freeze-dried magnetotactic bacteria (MSR-1) (carbon = 44%), the change in weight percentage as a function of temperature and the derivative of this change as a function of temperature when heated at a rate of 6 °C / min between 20 °C and 600 °C. (b) For a sample containing 3 mg of freeze-dried magnetotactic bacteria (MSR-1) (carbon = 44%), the heat flow (mW) generated by this sample as a function of temperature when heated at a rate of 6 °C / min between 20 °C and 600 °C. (c) For a sample containing 3 mg of freeze-dried magnetosomes (chains of magnetosomes, carbon = 7%) extracted from whole bacteria according to lysis condition 2 (condition n°2), the change in weight percentage as a function of temperature and the derivative of this change as a function of temperature when this sample is heated at a rate of 6 °C / min between 20 °C and 600 °. (d) For a sample containing 3 mg of freeze-dried magnetosomes (chains of magnetosomes, carbon = 7%) extracted from whole bacteria according to lysis condition 2 (condition n°2), the heat flow (mW) generated by this sample as a function of temperature when this sample is heated at a rate of 6 °C / min between 20 °C and 600 °. Regarding (a) and (c), the y-axis can be replaced by mass percentage to obtain the same curve.

[0014] Figure 2: TGA-DSC analysis of magnetosomes extracted from magnetotactic bacteria according to condition 3 and SIGMA nanoparticles synthesized chemically. (a) For a sample containing 3 mg of freeze-dried magnetosomes (magnetosomes, carbon = 4%) extracted from magnetotactic bacteria according to condition 3 (condition n°3), the change in weight percentage as a function of temperature and the derivative of this change as a function of temperature. (b) The heat flow (mW) generated by a sample containing 3 mg of freeze-dried magnetosomes extracted from magnetotactic bacteria according to condition 3 (condition n°3) (magnetosomes, carbon = 4%) as a function of temperature. (c) For a sample containing 3 mg of freeze-dried SIGMA nanoparticles (carbon = 0.3%), the change in weight percentage as a function of temperature and the derivative of this change as a function of temperature. (d) The heat flow (mW) generated by a sample containing 3 mg of freeze-dried SIGMA nanoparticles (carbon = 0.3%) as a function of temperature. For (a) and (c), the y-axis can be replaced by mass percentage to obtain the same curve.

[0015] Figure 3 : An illustrative example of the method according to the present invention can be used. The method according to the present invention utilizes pre-growth and growth stages to amplify magnetotactic bacteria and produce magnetosomes. The method of the present invention follows a series of pre-growth stages 1, i, and i + 1, where the pre-growth stage preferably starts by injecting nanoparticles-producing cells of the cell bank into a volume V PGS1 (usually about 50 ml), and the nanoparticles-producing cells are usually amplified in this volume for about 7 days, then the nanoparticles-producing cells are transferred from V PGS1 to V PGSi (V PGSi is usually about 500 ml), and the nanoparticles-producing cells are usually amplified in this volume for about 3 days, then the nanoparticles-producing cells are transferred from V PGSi to V PGSi+1 (V PGSi+1 is usually about 5 L), and the nanoparticles-producing cells are usually amplified in this volume for about 3 days until the optical density OD is usually equal to 1, then transferred to V GS0 (usually 45 liters), where, in the presence of oxygen bubbled through the growth medium, V GS0 is fed-batch cultured with a feed medium (usually 5 - 10 liters, per liter: ammonia (2.76.10 -1 M, 4.77 g); lactic acid (1.1 M, 100 g); K2HPO4 (1.72.10 -2 M, 3 g); MgSO4 (1.95.10 -3 M, 0.48 g); FeCl3 (7.4.10 -3 M, 2 g); thiamine (1.2.10 -6M, 0.0004 g); CaCl2 (1.3×10 -3 M, 0.2 g)) is supplemented to bring the optical density of bacterial growth to typically 5 to 40, and the amount of magnetosomes produced per liter of growth medium to 5 to 500 mg, where the pre-growth medium (per liter: NH4Cl (7.4×10 -3 M, 0.4 g); sodium lactate (2.3×10 -2 M, 2.6 g); K2HPO4 (5.74×10 -4 M, 0.1 g); MgSO4 (4.1×10 -4 M, 0.1 g); FeCl3 (1.8×10 -6 M, 0.0005 g); thiamine (1.2×10 -7 M, 0.00004 g); CaCl2 (0.9×10 -4 M, 0.015 g)) preferably contains a restricted concentration of iron or iron source (preferably <2 μM) to prevent nanoparticle production; the growth medium (per liter: NH4Cl (4.13×10 -3 M, 0.22 g); sodium lactate (1.4×10 -2 M, 1.3 g); K2HPO4 (1.55×10 -4 M, 0.027 g); MgSO4 (1.1×10 -4 M, 0.027 g); FeCl3 (1.8×10 -6 M, 0.0005 g); thiamine (1.2×10 -7 M, 0.00004 g); CaCl2 (0.9×10 -4 M, 0.015 g)) preferably contains iron or iron source (preferably >2 μM) to promote nanoparticle production. In the pre-growth stage: bacteria multiply abundantly and basically no nanoparticles are produced; in the growth stage: bacteria multiply abundantly and a large amount of nanoparticles are produced. In the pre-growth stage: air / oxygen bubbled into the growth medium: none or <200 mL air / O2 / minute, at the start of at least one pre-growth stage: 100 mbar < PO2 < 210 mbar (partial pressure of oxygen), at the end of at least one pre-growth stage: 0 < PO2 < 100 mbar. In the growth stage: air / oxygen bubbled into the growth medium: >200 mL air / O2 / minute, at the start of at least one growth stage: 100 mbar < PO2 < 210 mbar, at the end of at least one growth stage: 0 mbar < PO2 < 100 mbar.

[0016] Figure 4: An illustrative example of the pre-growth stage, showing the concentrations C1 and C2 of the carbon source, nitrogen source, and iron source at the start and end of the pre-growth stage. At the start of the pre-growth stage: C2 is approximately equal to 0 g / L; C 总量 is approximately equal to C1. At the end of the pre-growth stage: C1 < C2. C1 represents the content of carbon, nitrogen, and iron in the pre-growth medium without bacteria per liter of the pre-growth medium; C2 represents the content of carbon, nitrogen, and iron in the pre-growth medium with bacteria per liter of the pre-growth medium (note: C2 can also represent the quantity of carbon, nitrogen, and iron consumed by bacteria per liter of the pre-growth medium).

[0017] Figure 5 : An illustrative example of the growth stage (between T = 0 h and T = 20 h), showing the concentrations C1 and C2 of the carbon source, nitrogen source, and iron source at the start of the growth stage and 20 hours after the start. At the start of the growth stage (T = 0 h): C2 is approximately equal to 0 g / L; the total amount of C is approximately equal to C1. 20 hours after the start of the growth stage: C 1碳 ≥ C 2碳 ; C 1氮 ≥ C 2氮 ; C 1氮 = C 2氮 ; C 1氮 ≤ C 2氮 ; C 1铁 ≥ C 2铁 . C1 represents the content of carbon, nitrogen, and iron in the growth medium without bacteria per liter of the growth medium; C2 represents the content of carbon, nitrogen, and iron in the pre-growth medium with bacteria per liter of the growth medium (note: C2 can also represent the quantity of carbon, nitrogen, and iron consumed by bacteria per liter of the growth medium).

[0018] Figure 6 : An illustrative example of the growth stage (between T = 10 h and T = 40 h), showing the concentrations C1 and C2 of the carbon source, nitrogen source, and iron source 40 hours after the start of the growth stage and more than 40 hours after the start. 20 hours after the start of the growth stage: C 1碳 ≤ C 2碳 ; C 1氮 ≥ C 2氮 ; C 1氮 = C 2氮 ; C 1氮 ≤ C 2氮 ; C 1铁 ≤ C 2铁 ; More than 40 hours after the start of the growth stage: C 1碳 ≤ C 2碳 ; C 1氮 ≥ C 2氮 ; C 1氮 = C 2氮 ; C 1氮≤C 2氮 ; C 1铁 ≤C 2铁 。C1 represents the contents of carbon, nitrogen, and iron in the pre-growth medium without bacteria per liter of the growth medium; C2 represents the contents of carbon, nitrogen, and iron in the pre-growth medium with bacteria per liter of the growth medium. Detailed implementation mode

[0019] In one embodiment of the present invention, preferably, the nanoparticle-producing cells are preferably eukaryotic or prokaryotic cells that have the ability or function to produce or synthesize nanoparticles when they are injected into a medium in which the preferred concentration of at least one compound contained in the nanoparticles is greater than 10 -6 , 1 or 10 6 μM or when amplified in this medium.

[0020] In one embodiment of the present invention, the nanoparticle-producing cells basically do not produce nanoparticles, where these cells produce less than 10 50 , 10 10 , 10 5 , 10 3 , 10, 1, 10 -1 , 10 -3 or 10 -5 mg of nanoparticles, or preferably, the nanoparticle-containing cells in the pre-growth and / or growth and / or fed-batch medium contain less than 10 50 , 10 10 , 10 5 , 10 3 , 10, 1, 10 -1 , 10 -3 or 10 -5 mg of nanoparticles per liter of the pre-growth and / or growth and / or fed-batch medium.

[0021] In one embodiment of the present invention, the nanoparticle-producing cells produce nanoparticles, where these cells produce more than 0, 10 -50 , 10 -20 , 10 -10 , 10 -5 , 10 -1 , 1, 10, 10 3 or 10 5 mg of nanoparticles per liter of the pre-growth and / or growth and / or fed-batch medium, or preferably, the nanoparticle-containing cells in the pre-growth and / or growth and / or fed-batch medium contain less than 0, 10 -50 , 10 -20 , 10 -10 , 10 -5 , 10-1 1, 1, 10, 10 3 or 10 5 mg of nanoparticles per liter of pre-growth and / or growth and / or fed-batch medium.

[0022] In one embodiment of the present invention, the number of nanoparticles produced by nanoparticle cells during the pre-growth stage is equal to Q2 - Q1, where Q1 and Q2 are the numbers of nanoparticles produced at times t1 and t2 during the pre-growth stage, where t2 is greater than t1, preferably t2 / t1 is greater than 1, 2, 5, 10 or 10 3 , preferably t2 is the end time of the pre-growth stage, and t1 is the start time of the pre-growth stage.

[0023] In one embodiment of the present invention, the number of nanoparticles produced by nanoparticle cells during the growth stage is equal to Q2’ - Q1’, where Q1’ and Q2’ are the numbers of nanoparticles produced at times t1’ and t2’ during the growth stage, where t2’ is greater than t1’, preferably t2’ / t1’ is greater than 1, 2, 5, 10 or 10 3 , preferably t2’ is the end time of the growth stage, and t1’ is the start time of the growth stage.

[0024] Preferably Q2’ - Q1’ is greater than Q2 - Q1, preferably the multiple (factor) is at least 0, 10 -10 10 -1 1, 5, 10, 10 3 or 10 5 .

[0025] Preferably the total number of nanoparticles produced by nanoparticle-producing cells is equal to Q 总量 = Q2’ - Q1’ + Q2 - Q1.

[0026] In some cases, Q 总量 can be greater than 10 -50 10 -10 10 -5 10 -3 10 -1 0, 1, 5, 10 or 100 mg of nanoparticles, preferably greater than 10 -50 10 -10 10 -5 10 -3 10 -1 0, 1, 5, 10 or 100 mg of iron contained in the nanoparticles, preferably per liter of pre-growth and / or growth medium.

[0027] In some cases, Q 总量 can be less than 10 50, 10 10 , 10 5 , 10 3 , 100, 50, 10 or 1 mg of nanoparticles, preferably less than 10 50 , 10 10 , 10 5 , 10 3 , 100, 50, 10 or 1 mg of iron comprised in the nanoparticles, preferably per liter of pre-growth and / or growth medium.

[0028] In one embodiment of the invention, the yeast extract is an element selected from the group consisting of: i) a complete yeast extract, ii) a medium comprising more than 1, 10, 50 or 90% of compounds derived from a complete yeast extract, and iii) a chemical equivalent of a yeast extract.

[0029] In one embodiment of the invention, the peptone is an element selected from the group consisting of: i) a complete peptone, ii) a medium comprising more than 1, 10, 50 or 90% of compounds derived from a complete peptone, and iii) a chemical equivalent of a peptone.

[0030] The present invention relates to a method for producing high-purity iron oxide nanoparticles using nanoparticles to produce cell preparations, preferably comprising a pre-growth stage, comprising preferably amplifying nanoparticle-producing cells in a pre-growth medium, while preferably substantially not producing nanoparticles; preferably, followed by a growth stage, comprising preferably amplifying nanoparticle-producing cells from the pre-growth stage in a growth medium while producing nanoparticles, wherein the growth stage preferably differentiates itself from the pre-growth stage by at least one characteristic selected from the group consisting of:

[0031] i) The ratio of C FeGS / C FePGS is greater than 0, 10 -5 , 10 -3 , 1, 10, 10 3 or 10 5 , where C FeGS and C FePGS are the concentrations of iron or iron source in the growth medium and the pre-growth medium, respectively, ii) The ratio of C CGS / C CPGS is greater than 0, 10 -5 , 10 -3 , 1, 10, 10 3 or 10 5 , where C CGS and C CPGS are the concentrations of carbon or carbon source in the growth medium and the pre-growth medium, respectively, iii) CNGS / C NPGS has a ratio greater than 0.10 -5 , 10 -3 , 1, 10, 10 3 or 10 5 , where C NGS and C NPGS are the concentrations of nitrogen or nitrogen sources in the growth medium and the pre - growth medium, respectively, iv) ΔpH GS / ΔpH PGS has a ratio less than 0.10 10 , 10 5 , 10 3 , 10 2 , 1, 0.5 or 0.1, where ΔpH GS and ΔpH PGS are the changes in pH values in the growth medium and the pre - growth medium, respectively,

[0032] v) Q GGS / Q GPGS has a ratio greater than 0.10 10 , 10 5 , 10 3 , 10 2 , 1, 0.5 or 0.1, where Q GGS and Q GPGS are the amounts of gas, oxygen or air introduced or bubbled into the growth medium and the pre - growth medium, respectively,

[0033] vi) N SSGS / N SSPGS has a ratio less than 0.10 -5 , 10 -3 , 1, 10, 10 3 or 10 5 , where N SSGS and N SSPGS are the number of sub - stages in the growth phase and the number of sub - stages in the pre - growth phase, respectively, where the two sub - stages are separated by transferring the nanoparticle - producing cells from the first sub - stage to the second sub - stage, and

[0034] vii) the growth medium is supplemented with a fed - batch medium, while the pre - growth medium is not supplemented with a fed - batch medium, and / or

[0035] where, preferably, the pre - growth, growth and / or fed - batch medium do not include:

[0036] I) at least one compound or combination of compounds contained in or derived from yeast extract, where the at least one compound or combination of compounds is selected from the group consisting of:

[0037] I.1) More than 0, 10 -10 , 10 -5 , 10 -1 , 1, 10 3 or 10 5 grams of protein per liter of pre - growth and / or growth and / or fed - batch medium;

[0038] I.2) More than 0, 10 -10 , 10 -6 , 10 -1 , 1, 10 3 or 10 5 grams of nucleic acid per liter of pre - growth and / or growth and / or fed - batch medium;

[0039] I.3) More than 0, 10 -10 , 10 -5 , 10 -1 , 1, 10 3 or 10 5 grams of glutathione per liter of pre - growth and / or growth and / or fed - batch medium;

[0040] I.4) More than 0, 10 -10 , 10 -7 , 10 -1 , 1, 10 3 or 10 5 grams of at least one compound per liter of pre - growth and / or growth and / or fed - batch medium, where the at least one compound is selected from the group consisting of: dextran, mannan, trehalose, flavor nucleotides, B - vitamins, biotin, and volatile aromatic compounds,

[0041] I.5) More than 0, 10 -10 , 10 -7 , 10 -1 , 1, 10 3 or 10 5 grams of calcium per liter of pre - growth and / or growth and / or fed - batch medium,

[0042] I.6) More than 0, 10 -10 , 10 -6 , 10 -1 , 1, 10 3 or 10 5 grams of phosphorus per liter of pre - growth and / or growth and / or fed - batch medium,

[0043] I.7) More than 0, 10 -10 , 10 -8 , 10 -1 , 1, 10 3 or 105 0, 10

[0044] I.8) greater than 0, 10 -10 , 10 -1 , 1, 10 3 or 10 5 grams of chromium per liter of pre - growth and / or growth and / or fed - batch medium,

[0045] I.9) greater than 0, 10 -10 , 10 -7 , 10 -1 , 1, 10 3 or 10 5 grams of potassium per liter of pre - growth and / or growth and / or fed - batch medium,

[0046] I.10) greater than 0, 10 -10 , 10 -1 , 1, 10 3 or 10 5 grams of cobalt per liter of pre - growth and / or growth and / or fed - batch medium,

[0047] I.11) greater than 0, 10 -10 , 10 -9 , 10 -1 , 1, 10 3 or 10 5 grams of manganese per liter of pre - growth and / or growth and / or fed - batch medium,

[0048] I.12) greater than 0, 10 -10 , 10 -1 , 1, 10 3 or 10 5 grams of strontium per liter of pre - growth and / or growth and / or fed - batch medium,

[0049] I.13) greater than 0, 10 -10 , 10 -7 , 10 -1 , 1, 10 3 or 10 5 grams of magnesium per liter of pre - growth and / or growth and / or fed - batch medium,

[0050] I.14) greater than 0, 10 -10 , 10 -4 , 10 -1 , 1, 10 3 or 10 5 grams of yeast extract per liter of pre - growth and / or growth and / or fed - batch medium, and

[0051] I.15) More than 0, 1, 5, 10, 15, 20 or 50 different components derived from yeast extract;

[0052] II) At least one compound or combination of compounds contained in or derived from peptone, wherein at least one compound or combination of compounds is selected from the group consisting of:

[0053] II.1) More than 0, 10 -10 , 10 -4 , 10 -1 , 1, 10 3 or 10 5 grams of ash per liter of pre-growth and / or growth and / or fed-batch medium,

[0054] II.2) More than 0, 10 -10 , 10 -3 , 10 -1 , 1, 10 3 or 10 5 grams of protein per liter of pre-growth and / or growth and / or fed-batch medium,

[0055] II.3) More than 0, 10 -10 , 10 -4 , 10 -1 , 1, 10 3 or 10 5 grams of sucrose per liter of pre-growth and / or growth and / or fed-batch medium,

[0056] II.4) More than 0, 10 -10 , 10 -5 , 10 -1 , 1, 10 3 or 10 5 grams of raffinose per liter of pre-growth and / or growth and / or fed-batch medium,

[0057] II.5) More than 0, 10 -10 , 10 -4 , 10 -1 , 1, 10 3 or 10 5 grams of neutral detergent fiber per liter of pre-growth and / or growth and / or fed-batch medium,

[0058] II.6) More than 0, 10 -10 , 10 -5 , 10 -1 , 1, 10 3 or 10 5 grams of ether extract per liter of pre-growth and / or growth and / or fed-batch medium,

[0059] II.7) greater than 0, 10 -10 , 10 -5 , 10 -1 , 1, 10 3 or 10 5 grams of peptone per liter of pre - growth and / or growth and / or fed - batch medium, and

[0060] II.8) greater than 0, 1, 5, 10, 15, 20 or 50 different components from peptone; III) comprising at least one compound or combination of compounds from Wolfe's minerals or mineral elixirs, wherein at least one compound or combination of compounds is selected from the group consisting of:

[0061] III.1) greater than 0, 10 -20 , 10 -5 , 10 -1 , 1, 10 3 or 10 5 moles of nitrilotriacetic acid per liter of pre - growth and / or growth and / or fed - batch medium,

[0062] III.2) greater than 0, 10 -20 , 10 -6 , 10 -1 , 1, 10 3 or 10 5 moles of magnesium sulfate per liter of pre - growth and / or growth and / or fed - batch medium,

[0063] III.3) greater than 0, 10 -20 , 10 -5 , 10 -1 , 1, 10 3 or 10 5 moles of sodium chloride per liter of pre - growth and / or growth and / or fed - batch medium,

[0064] III.4) greater than 0, 10 -20 , 10 -5 , 10 -1 , 1, 10 3 or 10 5 moles of manganese sulfate per liter of pre - growth and / or growth and / or fed - batch medium,

[0065] III.5) greater than 0, 10 -20 , 10 -6 , 10 -1 , 1, 10 3 or 10 50.1 molar of ferrous sulfate per liter of pre-growth and / or growth and / or fed-batch medium,

[0066] III.6) Above 0, 0.1 -20 、0.1 -6 、0.1 -1 、1、0.1 3 or 0.1 5 molar of cobalt nitrate per liter of pre-growth and / or growth and / or fed-batch medium,

[0067] III.7) Above 0, 0.1 -20 、0.1 -6 、0.1 -1 、1、0.1 3 or 0.1 5 molar of calcium chloride per liter of pre-growth and / or growth and / or fed-batch medium,

[0068] III.8) Above 0, 0.1 -20 、0.1 -6 、0.1 -1 、1、0.1 3 or 0.1 5 molar of zinc sulfate per liter of pre-growth and / or growth and / or fed-batch medium,

[0069] III.9) Above 0, 0.1 -20 、0.1 -7 、0.1 -1 、1、0.1 3 or 0.1 5 molar of copper sulfate per liter of pre-growth and / or growth and / or fed-batch medium,

[0070] III.10) Above 0, 0.1 -20 、0.1 -7 、0.1 -1 、1、0.1 3 or 0.1 5 molar of potassium alum from Wolff's minerals or mineral elixirs per liter of pre-growth and / or growth and / or fed-batch medium,

[0071] III.11) Above 0, 0.1 -20 、0.1 -7 、0.1 -1 、1、0.1 3 or 0.1 5 molar of boric acid per liter of pre-growth and / or growth and / or fed-batch medium,

[0072] III.12) Above 0, 0.1 -20 、0.1 -8 、0.1 -1 、1、0.13 or 10 5 moles of sodium molybdate per liter of pre - growth and / or growth and / or fed - batch medium,

[0073] III.13) greater than 0, 10 -20 、10 -9 、10 -1 、1、10 3 or 10 5 moles of sodium selenite per liter of pre - growth and / or growth and / or fed - batch medium,

[0074] III.14) greater than 0, 10 -20 、10 -7 、10 -1 、1、10 3 or 10 5 moles of sodium tungstate per liter of pre - growth and / or growth and / or fed - batch medium,

[0075] III.15) greater than 0, 10 -20 、10 -7 、10 -1 、1、10 3 or 10 5 moles of nickel chloride per liter of pre - growth and / or growth and / or fed - batch medium,

[0076] III.16) greater than 0, 10 -20 、10 -8 、10 -1 、1、10 3 or 10 5 moles of Wolfe's minerals or mineral elixirs per liter of pre - growth and / or growth and / or fed - batch medium, and

[0077] III.17) greater than 0, 10 different components from Wolfe's minerals or mineral elixirs; IV) greater than 0, 10 -20 、10 -5 、10 -1 、1、10 3 or 10 5 moles of EDTA per liter of pre - growth and / or growth and / or fed - batch medium;

[0078] and / or

[0079] V) at least one compound or combination of compounds contained in or derived from Wolfe's vitamins, wherein the at least one compound or combination of compounds is selected from the group consisting of: V.1) greater than 0, 10 -20 、10 -9 、10 -1 、1、103 or 10 5 moles of biotin per liter of pre - growth and / or growth and / or fed - batch medium,

[0080] V.2) greater than 0, 10 -20 、10 -8 、10 -1 、1、10 3 or 10 5 moles of calcium pantothenate per liter of pre - growth and / or growth and / or fed - batch medium,

[0081] V.3) greater than 0, 10 -20 、10 -9 、10 -1 、1、10 3 or 10 5 moles of folic acid per liter of pre - growth and / or growth and / or fed - batch medium,

[0082] V.4) greater than 0, 10 -20 、10 -9 、10 -1 、1、10 3 or 10 5 moles of inositol per liter of pre - growth and / or growth and / or fed - batch medium,

[0083] V.5) greater than 0, 10 -20 、10 -8 、10 -1 、1、10 3 or 10 5 moles of niacin per liter of pre - growth and / or growth and / or fed - batch medium,

[0084] V.6) greater than 0, 10 -20 、10 -8 、10 -1 、1、10 3 or 10 5 moles of p - aminobenzoic acid per liter of pre - growth and / or growth and / or fed - batch medium,

[0085] V.7) greater than 0, 10 -20 、10 -9 、10 -1 、1、10 3 or 10 5 moles of pyridoxine hydrochloride per liter of pre - growth and / or growth and / or fed - batch medium,

[0086] V.8) greater than 0, 10 -20 、10 -9 、10 -1 、1、103 or 10 5 moles of riboflavin per liter of pre - growth and / or growth and / or fed - batch medium,

[0087] V.9) greater than 0, 10 -20 、10 -9 、10 -1 、1、10 3 or 10 5 moles of thiamine hydrochloride per liter of pre - growth and / or growth and / or fed - batch medium,

[0088] V.10) greater than 0, 10 -20 、10 -8 、10 -1 、1、10 3 or 10 5 moles of lipoic acid per liter of pre - growth and / or growth and / or fed - batch medium,

[0089] V.11) greater than 0, 10 -20 、10 -9 、10 -1 、1、10 3 or 10 5 moles of at least one component of the Wolfe vitamins, and

[0090] V.12) greater than 0, 1, 5, 10, 15 or 20 different Wolfe vitamins; and / or

[0091] Preferably, where measured per milligram or per gram or per milliliter or per liter of pre - growth, growth and / or fed - batch medium / media, the pre - growth, growth and / or fed - batch medium preferably does not contain more than:

[0092] 1) 0, 10 -20 、10 -10 、10 -5 、10 -3 or 1 gram of protein preferably derived from yeast extract,

[0093] 2) 0, 10 -20 、10 -10 、10 -6 、10 -3 or 1 gram of nucleic acid preferably derived from yeast extract,

[0094] 3) 0, 10 -20 、10 -10 、10 -5 、10 -3 or 1 gram of glutathione preferably derived from yeast extract,

[0095] 4) 0, 10-20 、 10 -10 、 10 -7 、 10 -1 or 1 g of at least one compound preferably derived from yeast extract, wherein the at least one compound is selected from the group consisting of: glucan, mannan, trehalose, flavor nucleotides, B vitamins, biotin, and volatile aromatic compounds,

[0096] 5) 0, 10 -20 、 10 -10 、 10 -7 、 10 -1 or 1 g of calcium preferably derived from yeast extract,

[0097] 6) 0, 10 -20 、 10 -10 、 10 -6 、 10 -1 or 1 g of phosphorus preferably derived from yeast extract,

[0098] 7) 0, 10 -20 、 10 -10 、 10 -8 、 10 -1 or 1 g of zinc preferably derived from yeast extract,

[0099] 8) 0, 10 -20 、 10 -10 、 10 -5 、 10 -1 or 1 g of chromium preferably derived from yeast extract,

[0100] 9) 0, 10 -20 、 10 -10 、 10 -7 、 10 -5 、 10 -1 or 1 g of potassium preferably derived from yeast extract,

[0101] 10) 0, 10 -20 、 10 -10 、 10 -5 、 10 -1 or 1 g of cobalt preferably derived from yeast extract,

[0102] 11) 0, 10 -20 、 10 -9 、 10 -5 、 10 -1 or 1 g of manganese preferably derived from yeast extract,

[0103] 12) 0, 10 -20 、 10 -10 、 10 -5 、 10-1 or 1 g of strontium preferably derived from yeast extract,

[0104] 13) 0, 10 -20 , 10 -10 , 10 -7 , 10 -3 , 10 -1 or 1 g of magnesium preferably derived from yeast extract,

[0105] 14) 0, 10 -20 , 10 -10 , 10 -4 , 10 -1 or 1 g of yeast extract,

[0106] 15) 0, 1, 5, 10 or 15 different components from yeast extract,

[0107] 16) 0, 10 -20 , 10 -10 , 10 -4 , 10 -1 or 1 g of ash preferably derived from peptone,

[0108] 17) 0, 10 -20 , 10 -10 , 10 -3 , 10 -1 or 1 g of protein preferably derived from peptone,

[0109] 18) 0, 10 -20 , 10 -10 , 10 -4 , 10 -1 or 1 g of sucrose preferably derived from peptone,

[0110] 19) 0, 10 -20 , 10 -10 , 10 -5 , 10 -1 or 1 g of raffinose preferably derived from peptone,

[0111] 20) 0, 10 -20 , 10 -10 , 10 -4 , 10 -1 or 1 g of neutral detergent fiber preferably derived from peptone,

[0112] 21) 0, 10 -20 , 10 -10 , 10 -5 , 10 -1 or 1 g of ether extract preferably derived from peptone,

[0113] 22) 0, 10 -20 , 10 -10 , 10 -5 , 10 -1 or 1 g of peptone,

[0114] 23) 0, 1, 5, 7, 10 or 15 different components from peptone,

[0115] 24) 0, 10 -20 , 10 -10 , 10 -5 , 10 -1 or 1 mol of nitrilotriacetic acid preferably sourced from Wolff minerals or mineral elixirs,

[0116] 25) 0, 10 -20 , 10 -10 , 10 -6 , 10 -1 or 1 mol of magnesium sulfate preferably sourced from Wolff minerals or mineral elixirs,

[0117] 26) 0, 10 -20 , 10 -10 , 10 -5 , 10 -1 or 1 mol of sodium chloride preferably sourced from Wolff minerals or mineral elixirs,

[0118] 27) 0, 10 -20 , 10 -10 , 10 -5 , 10 -1 or 1 mol of manganese sulfate preferably sourced from Wolff minerals or mineral elixirs,

[0119] 28) 0, 10 -20 , 10 -10 , 10 -6 , 10 -1 or 1 mol of ferrous sulfate preferably sourced from Wolff minerals or mineral elixirs,

[0120] 29) 0, 10 -20 , 10 -10 , 10 -6 , 10 -1 or 1 mol of cobalt nitrate preferably sourced from Wolff minerals or mineral elixirs,

[0121] 30) 0, 10 -20 , 10 -10 , 10 -6 , 10 -1 or 1 mol of calcium chloride preferably sourced from Wolff minerals or mineral elixirs,

[0122] 31) 0, 10 -20 , 10 -10 , 10 -6 , 10 -1 or 1 mol of zinc sulfate preferably sourced from Wolff minerals or mineral elixirs,

[0123] 32) 0, 10 -20 , 10 -10 , 10 -7 , 10 -1 or 1 mol of copper sulfate preferably sourced from Wolff minerals or mineral elixirs,

[0124] 33) 0, 10 -20 , 10 -10 , 10 -7 , 10 -1 or 1 mol of potassium alum preferably sourced from Wolff minerals or mineral elixirs,

[0125] 34) 0, 10 -20 , 10 -10 , 10 -7 , 10 -1 or 1 mol of boric acid preferably sourced from Wolff minerals or mineral elixirs,

[0126] 35) 0, 10 -20 , 10 -10 , 10 -8 , 10 -3 , 10 -1 or 1 mol of sodium molybdate preferably sourced from Wolff minerals or mineral elixirs,

[0127] 36) 0, 10 -20 , 10 -15 , 10 -9 , 10 -3 , 10 -1 or 1 mol of sodium selenite preferably sourced from Wolff minerals or mineral elixirs,

[0128] 37) 0, 10 -20 , 10 -7 , 10 -3 , 10 -1 or 1 mol of sodium tungstate preferably sourced from Wolff minerals or mineral elixirs,

[0129] 38) 0, 10 -20 , 10 -7 , 10 -3 , 10 -1 or 1 mol of nickel chloride preferably sourced from Wolff minerals or mineral elixirs,

[0130] 39) 0, 10-20 , 10 -8 , 10 -3 , 10 -1 or 1 mol of Wolff minerals or mineral elixirs,

[0131] 40) 0, 1, 5, 10 or 20 different components of Wolff minerals or mineral elixirs,

[0132] 41) 0, 10 -20 , 10 -10 , 10 -5 , 10 -1 or 1 mol of EDTA,

[0133] 42) 0, 10 -50 , 10 -30 , 10 -9 , 10 -3 , 10 -1 or 1 mol of biotin preferably derived from Wolff vitamins,

[0134] 43) 0, 10 -50 , 10 -30 , 10 -8 , 10 -3 , 10 -1 or 1 mol of calcium pantothenate preferably derived from Wolff vitamins,

[0135] 44) 0, 10 -50 , 10 -30 , 10 -9 , 10 -3 , 10 -1 or 1 mol of folic acid preferably derived from Wolff vitamins,

[0136] 45) 0, 10 -50 , 10 -30 , 10 -9 , 10 -3 , 10 -1 or 1 mol of inositol preferably derived from Wolff vitamins,

[0137] 46) 0, 10 -50 , 10 -30 , 10 -10 , 10 -8 , 10 -3 , 10 -1 or 1 mol of nicotinic acid preferably derived from Wolff vitamins,

[0138] 47) 0, 10 -50 , 10 -30 , 10 -8 , 10 -3, 10 -1 or 1 mol of p-aminobenzoic acid preferably sourced from Wolff Vitamins,

[0139] 48) 0, 10 -50 , 10 -30 , 10 -9 , 10 -3 , 10 -1 or 1 mol of pyridoxine hydrochloride preferably sourced from Wolff Vitamins,

[0140] 49) 0, 10 -50 , 10 -30 , 10 -9 , 10 -3 , 10 -1 or 1 mol of riboflavin preferably sourced from Wolff Vitamins,

[0141] 50) 0, 10 -50 , 10 -30 , 10 -9 , 10 -3 , 10 -1 or 1 mol of thiamine hydrochloride preferably sourced from Wolff Vitamins,

[0142] 51) 0, 10 -50 , 10 -30 , 10 -8 , 10 -3 , 10 -1 or 1 mol of lipoic acid preferably sourced from Wolff Vitamins,

[0143] 52) 0, 10 -50 , 10 -30 , 10 -9 , 10 -3 , 10 -1 or 1 mol of components of at least one Wolff Vitamin,

[0144] 53) 0, 1, 5, 10 or 20 different Wolff Vitamins,

[0145] 54) 0, 1, 2, 3, 6, 10 or 100 different vitamins,

[0146] 55) 0, 10 -50 , 10 -20 , 10 -9 , 10 -8 , 10 -7 , 10 -5 , 10 -3 , 10 -1 or 1 mol of at least one vitamin,

[0147] 56) 0, 10 -50 , 10 -20 , 10 -10 , 10 -5 , 10 -4 , 10 -2 , 10 -1 , 1, 10 or 10 3 g of yeast extract,

[0148] 57) 0, 10 -50 , 10 -20 , 10 -9 , 10 -3 , 10 -1 , 1, 5 or 10 mol of components of at least one yeast extract,

[0149] 58) 0, 1, 2, 5, 10 or 100 different components of yeast extract,

[0150] 59) 0, 10 -50 , 10 -10 , 10 -5 , 10 -1 , 1, 10 or 10 3 g of peptone,

[0151] 60) 0, 10 -50 , 10 -9 , 10 -5 , 10 -3 , 10 -1 , 1 or 10 mol of components of at least one peptone,

[0152] 61) 0, 1, 2, 5, 10 or 100 different components of peptone,

[0153] 62) 0, 1, 2, 5, 10 or 100 different CMR reagents,

[0154] 63) 0, 10 -50 , 10 -9 , 10 -5 , 0.05, 10 -1 , 1, 10, 10 3 or 10 6 mg of at least one CMR reagent,

[0155] 64) 0, 1, 2, 5, 10 or 100 different chelating agents,

[0156] 65) 0, 10 -50 , 10 -20 , 10 -9 , 10 -8 , 10 -3 , 10-1 1, 5, 10 or 10 3 mol of at least one chelating agent

[0157] 66) 0, 1, 2, 5, 10 or 100 different amino acids

[0158] 67) 0, 10 -50 10 -10 10 -5 10 -3 1, 10, 10 3 10 5 or 10 10 mg of at least one amino acid

[0159] 68) 0, 1, 2, 5, 10 or 100 different toxic or cytotoxic compounds

[0160] 69) 0, 10 -50 10 -10 10 -5 10 -3 10 -1 1, 10, 10 3 or 10 5 mg of at least one toxic or cytotoxic compound

[0161] 70) 0, 1, 3 or 7 heavy metals different from iron

[0162] 71) 0, 10 -50 10 -10 10 -5 10 -3 10 -1 1, 10, 10 3 or 10 5 mg of at least one heavy metal different from iron

[0163] 72) 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 metals or chemical elements selected from cadmium, lead, arsenic, mercury, cobalt, vanadium, nickel, lithium, antimony and copper

[0164] 73) 1 mg of cadmium, lead, arsenic, mercury, cobalt, vanadium, nickel, lithium, antimony and / or copper

[0165] 74) 0, 10 -50 10 -10 0.5, 1, 5, 10, 10 3 or 10 6 ml or 10 -50 10 -30 10 -10 10 -8 10-5 , 10 -3 , 1, 10, 10 3 or 10 6 mol of Wolf vitamins,

[0166] 75) 10 -50 , 10 -20 , 10 -9 , 10 -5 , 10 -1 , 1, 5, 10, 10 3 or 10 5 mol of at least one Wolf vitamin component,

[0167] 76) 0, 1, 2, 5, 10 or 100 different components of Wolf vitamins,

[0168] 77) 0, 10 -50 , 10 -10 , 0.5, 1, 5, 10, 10 3 or 10 6 mL or 10 -50 , 10 -30 , 10 -10 , 10 -8 , 10 -5 , 10 -3 , 1, 10, 10 3 or 10 6 mol of Wolf minerals,

[0169] 78) 10 -50 , 10 -20 , 10 -9 , 10 -5 , 10 -1 , 1, 5, 10, 10 3 or 10 5 mol of at least one Wolf mineral component,

[0170] 79) 0, 1, 2, 5, 7, 10, 15 or 100 different components of Wolf minerals,

[0171] 80) 10 -50 , 10 -20 , 10 -9 , 10 -5 , 10 -1 , 1, 5, 10, 10 3 or 10 5 mol of mineral elixirs,

[0172] 81) 10 -50 , 10 -20 , 10 -9, 10 -5 , 10 -1 , 1, 5, 10, 10 3 or 10 5 mol of the components of at least one mineral elixir, and / or

[0173] 82) 0, 1, 2, 5, 10, 14 or 100 different components of mineral elixirs.

[0174] The present invention also relates to a method according to the present invention, wherein the fed-batch culture medium contains at least one compound selected from the group consisting of iron, iron source, carbon, carbon source, nitrogen, nitrogen source and combinations thereof, and the concentration of at least one compound in the fed-batch culture medium is greater than 10 -6 μM, 1 μM and / or 10 6 μM.

[0175] In one embodiment of the present invention, at least one vitamin component of the Wolfe vitamin or vitamin solution is selected from the group consisting of folic acid, folate, pyridoxol, pyridoxamine, pyridoxal, riboflavin, biotin, thiamine, niacin, pantothenic acid, vitamin B12, aminobenzoic acid, lipoic acid, all-trans retinol, retinal, alternative provitamin A-functionalized carotenoids including all-trans β-carotene, nicotinic acid, nicotinamide, nicotinamide riboside, cyanocobalamin, hydroxocobalamin, methylcobalamin, adenosylcobalamin, ascorbic acid, cholecalciferol, ergocalciferol, tocopherol, tocotrienol, menadione, phylloquinone, vitamin A, vitamin B1, vitamin vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin B 12 , vitamin C, vitamin D, vitamin D2, vitamin D3, vitamin E, vitamin K, vitamin V i (where V can be any letter from A to Z and i can be any integer between 1 and 100), and their derivatives.

[0176] In another embodiment of the present invention, at least one component of the Wolfe minerals is selected from the group consisting of nitrilotriacetic acid, magnesium sulfate, sodium chloride, manganese sulfate, ferrous sulfate heptahydrate, cobalt nitrate, calcium chloride, zinc sulfate heptahydrate, copper sulfate pentahydrate, potassium alum dodecahydrate, boric acid, sodium molybdate, sodium selenite, sodium tungstate dihydrate, nickel chloride and their derivatives.

[0177] In another embodiment of the present invention, at least one component of the yeast extract is selected from the group consisting of: i) at least one protein, ii) at least one nucleic acid, iii) at least one functional peptide, iv) glutathione, v) glucan, vi) mannan, vii) trehalose, viii) flavor nucleotides, ix) B vitamins, x) biotin, x) at least one volatile aromatic compound, xi) calcium, xii) phosphorus, xiii) zinc, xiv) iron, xv) chromium, xvi) potassium, xvii) cobalt, xviii) manganese, xix) strontium, xx) magnesium, and xxi) their derivatives.

[0178] In another embodiment of the present invention, at least one component of the mineral elixir is selected from the group consisting of nitrilotriacetic acid, MgSO4, MnSO4, NaCl, FeSO4, CoSO4, CaCl2, ZnSO4, CuSO4, KAl(SO4)2, H3BO3, Na2MoO4, NiCl2, Na2SeO3, and their derivatives.

[0179] In one embodiment of the present invention, the following two statements are equivalent: the pre-growth, growth, and / or fed-batch medium does not contain more than a certain amount, concentration, or quantity of at least one compound and the pre-growth, growth, and / or fed-batch medium contains less than a certain amount, concentration, or quantity of at least one compound.

[0180] In one embodiment of the present invention, the nanoparticle-producing cells are expanded or grown in the pre-growth and / or growth and / or fed-batch medium, preferably expanded or grown in the pre-growth and / or growth medium rather than in the fed-batch medium.

[0181] In one embodiment of the present invention, the growth and / or pre-growth medium contains the growth and / or pre-growth medium: i) without the need to inject the fed-batch medium into the growth and / or pre-growth medium or before injecting the fed-batch medium into the growth and / or pre-growth medium; or ii) with the need to inject the fed-batch medium into the growth and / or pre-growth medium or after injecting the fed-batch medium into the growth and / or pre-growth medium.

[0182] In one embodiment of the present invention, the fed-batch medium is the fed-batch medium or the medium before it is injected into the growth and / or pre-growth medium.

[0183] In another embodiment of the present invention, the fed-batch medium is a part of the pre-growth and / or pre-growth medium after it is injected into the growth and / or pre-growth medium.

[0184] In one embodiment of the present invention, parameter C FeGS 、CFePGS 、 C CGS 、 C CPGS 、 C NGS 、 C NPGS 、 ΔpH GS and / or ΔpH PGS are present in or measured in the pre-growth, growth and / or fed-batch medium.

[0185] In one embodiment of the invention, the parameters Q GGS 、 Q GPGS 、 N SSGS and N SSPGS and the case where the growth medium is supplemented with the fed-batch medium while the pre-growth medium is not supplemented with the fed-batch medium are measured at the start or end of the pre-growth and / or growth phase.

[0186] In one embodiment of the invention, the derivative is a derivative of at least one first compound selected from the group consisting of: i) a second compound that differs from the at least one first compound by at least one different atom or functional group, preferably a second compound derived from the at least one first compound after transformation of the at least one first compound, preferably the second compound and the at least one first compound having at least one common atom or functional group, ii) a non-hydrated form of the at least one first compound, iii) a hydrated form of the at least one first compound, iv) a reduced form of the at least one first compound, v) an oxidized form of the at least one first compound, vi) an acidic form of the at least one first compound, v) a basic form of the at least one first compound, vi) a crystalline or solid form of the at least one first compound, vii) a soluble or dissolved form of the at least one first compound, and x) a salt of the at least one first compound.

[0187] The present invention relates to a method according to the present invention, wherein the growth phase differentiates itself from the pre-growth phase by at least one characteristic selected from the group consisting of:

[0188] i) The ratio of C FeGS / C FePGS is greater than 10 -5 、 10 -3 、 1、 10、 10 3 or 10 5 where C FeGS and C FePGS are the concentrations of iron or iron source in the growth medium and the pre-growth medium, respectively, ii) The ratio of C CGS / C CPGS is greater than 10 -5 、 10 -3 、 1、 10、 10 3 or 105 , where C CGS and C CPGS are the concentrations of carbon or carbon sources in the growth medium and the pre-growth medium, respectively, iii) the ratio of C NGS / C NPGS is greater than 10 -5 , 10 -3 , 1, 10, 10 3 or 10 5 , where C NGS and C NPGS are the concentrations of nitrogen or nitrogen sources in the growth medium and the pre-growth medium, respectively, iv) the ratio of ΔpH GS / ΔpH PGS is less than 10 10 , 10 5 , 10 3 , 10 2 , 1, 0.5 or 0.1, where ΔpH GS and / ΔpH PGS are the changes in pH values in the growth medium and the pre-growth medium, respectively,

[0189] v) the ratio of Q GGS / Q GPGS is greater than 10 10 , 10 5 , 10 3 , 10 2 , 1, 0.5 or 0.1, where Q GGS and Q GPGS are the amounts of gas, oxygen or air introduced or bubbled into the growth medium and the pre-growth medium, respectively,

[0190] vi) the ratio of N SSGS / N SSPGS is less than 10 -5 , 10 -3 , 1, 10, 10 3 or 10 5 , where N SSGS and N SSPGS are the number of sub-stages in the growth phase and the number of sub-stages in the pre-growth phase, respectively, where the two sub-stages are separated by transferring the nanoparticle-producing cells from the first sub-stage (preferably associated with the growth of nanoparticle-producing cells in the first volume) to the second sub-stage (preferably associated with the growth of nanoparticle-producing cells in the second volume), and

[0191] vii) the growth medium is supplemented with a fed-batch medium, while the pre-growth medium is not supplemented with a fed-batch medium.

[0192] The present invention also relates to a method according to the present invention, wherein the pre-growth and / or growth and / or fed-batch culture medium preferably contains less than per kilogram or per liter of the pre-growth and / or growth and / or fed-batch culture medium:

[0193] i) 10 by mass or volume 3 %, or 1% by mass or volume, or 10 by mass or volume -2 %, or 5·10 by mass or volume -3 %, or 10 3 g, or 10 2 g, or 10 g, or 10 10 mL, or 10 5 mL, or 10 3 mL, or 10 mL, or 5 mL, or 1 mL, or 0.5 mL, or 10 -5 mL, or 10 3 mol, or 10 mol, or 1 mol, or 10 -5 mol, or 10 -8 mol, or 10 -9 mol, or 10 -10 mol of vitamins selected from the group consisting of, preferably Wolfe vitamins, or chemical components: folic acid, folate, pyridoxine, pyridoxine hydrochloride, pyridoxamine, pyridoxal, riboflavin, biotin, thiamine, thiamine hydrochloride, niacin, pantothenic acid, calcium pantothenate, inositol, p-aminobenzoic acid, aminobenzoic acid, lipoic acid, all-trans retinol, retinal, alternative provitamin A-functionalized carotenoids including all-trans β-carotene, nicotinic acid, nicotinamide, nicotinamide, nucleosides, cyanocobalamin, hydroxocobalamin, methylcobalamin, adenosylcobalamin, ascorbic acid, cholecalciferol, ergocalciferol, tocopherols, tocotrienols, menadione, phylloquinone, vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin B 12 , vitamin C, vitamin D, vitamin D2, vitamin D3, vitamin E, vitamin K, vitamin V i and their derivatives, where V can be any letter from A to Z, and i can be any integer from 1 to 100,

[0194] ii) 1, 5, 6, 10 or 20 different vitamins selected from the group consisting of, preferably Wolf vitamins, or chemical components: folic acid, folate, pyridoxine, pyridoxine hydrochloride, pyridoxamine, pyridoxal, riboflavin, biotin, thiamine, thiamine hydrochloride, niacin, pantothenic acid, calcium pantothenate, inositol, p-aminobenzoic acid, aminobenzoic acid, lipoic acid, all-trans retinol, retinal, alternative provitamin A-functionalized carotenoids including all-trans β-carotene, nicotinic acid, nicotinamide, nicotinamide, nucleosides, cyanocobalamin, hydroxocobalamin, methylcobalamin, adenosylcobalamin, ascorbic acid, cholecalciferol, ergocalciferol, tocopherols, tocotrienols, menadione, phylloquinone, vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin B 12 , vitamin C, vitamin D, vitamin D2, vitamin D3, vitamin E, vitamin K, vitamin V i and their derivatives, where V can be any letter from A to Z, and i can be any integer from 1 to 100,

[0195] iii) 10 3 % by mass or volume, or 10% by mass or volume, or 1% by mass or volume, or 10 -2 % by mass or volume, or 10 -5 % or 10 3 g, or 10 g, or 1 g, or 10 5 mL, or 10 3 mL, or 10 mL, or 1 mL, or 10 -3 mL, or 10 3 mol, or 10 mol, or 1 mol, or 10 -3 mol, or 10 -7 mol, or 10 -8 mol, or 10 - 10 mol of minerals selected from the group consisting of, preferably Wolf minerals or mineral elixirs, or chemical components: nitrilotriacetic acid, magnesium sulfate, sodium chloride, manganese sulfate, ferrous sulfate, ferrous sulfate heptahydrate, cobalt nitrate, calcium chloride, zinc sulfate, zinc sulfate heptahydrate, copper sulfate, copper sulfate pentahydrate, potassium alum, potassium alum dodecahydrate, boric acid, sodium molybdate, sodium selenite, sodium tungstate, sodium tungstate dihydrate, nickel chloride, EDTA, MgSO4, MnSO4, NaCl, FeSO4, CoSO4, CaCl2, ZnSO4, CuSO4, KAl(SO4)2, H3BO3, Na2MoO4, NiCl2, Na2SeO3 and their derivatives,

[0196] iv) Minerals selected from the group consisting of 1, 3, 7, or 10 different components, preferably Wolff minerals or mineral elixirs, or chemical components: nitrilotriacetic acid, magnesium sulfate, sodium chloride, manganese sulfate, ferrous sulfate, ferrous sulfate heptahydrate, cobalt nitrate, calcium chloride, zinc sulfate, zinc sulfate heptahydrate, copper sulfate, copper sulfate pentahydrate, potassium alum, potassium alum dodecahydrate, boric acid, sodium molybdate, sodium selenite, sodium tungstate, sodium tungstate dihydrate, nickel chloride, EDTA, MgSO4, MnSO4, NaCl, FeSO4, CoSO4, CaCl2, ZnSO4, CuSO4, KAl(SO4)2, H3BO3, Na2MoO4, NiCl2, Na2SeO3, and their derivatives,

[0197] v) 10 -50 g, or 10 -10 g, or 10 -5 g, or 0.005 g, or 10 -1 g, or 1 g, or 10 g or 10 5 g, or 10 -50 M, or 10 -8 M, or 10 -3 M, or 10 -1 M, or 1 M or 10 3 Components of at least one yeast extract or at least one compound derived from the group of yeast extracts, selected from the group consisting of: at least one protein, at least one nucleic acid, at least one functional peptide, glutathione, glucan, mannan, trehalose, flavoring nucleotide, B vitamins, biotin, at least one volatile aromatic compound, calcium, phosphorus, zinc, iron, chromium, potassium, cobalt, manganese, strontium, magnesium, and their derivatives,

[0198] vi) 1, 2, 3, 5, 10, 15, 20, or 50 different components of yeast extracts or different compounds derived from yeast extracts, selected from the group consisting of: at least one protein, at least one nucleic acid, at least one functional peptide, glutathione, glucan, mannan, trehalose, flavoring nucleotides, B vitamins, biotin, at least one volatile aromatic compound, calcium, phosphorus, zinc, iron, chromium, potassium, cobalt, manganese, strontium, magnesium, and their derivatives,

[0199] vii) 10 -50 g, or 10 -10 g, or 10 -3 g, or 0.01 g, or 1 g, or 5 g, or 10 g or 10 5 g, or 10 -50 M, or 10 - 20 M, or 10 -8M, or 10 -3 M, or 10 -1 M, or 1M, or 10M or 10 3 at least one peptone component or at least one compound derived from peptone, selected from the group consisting of: ash, protein, sucrose, stachyose, raffinose, neutral detergent fiber, ethereal extract, and derivatives thereof

[0200] viii) 1, 3, 5, 10, 20 or 50 different components of peptone or different components of compounds derived from peptone, selected from the group consisting of: ash, protein, sucrose, stachyose, raffinose, neutral detergent fiber, ethereal extract, and derivatives thereof

[0201] ix) 10 -50 , 10 -10 , 10 -5 , 0.001, 10 -1 , 1, 10, 10 3 or 10 5 grams of EDTA

[0202] x) 10 -50 , 10 -10 , 10 -5 , 0.001, 10 -1 , 1, 10, 10 3 or 10 5 grams of at least one amino acid

[0203] xi) 1, 3, 5, 10, 20 or 50 different amino acids

[0204] xii) 1, 5, 7, 12, 15, 20 or 50 different CMR, toxic or cytotoxic compounds, selected from the group consisting of: nitrilotriacetic acid, manganese sulfate, cobalt nitrate, zinc sulfate, copper sulfate, potassium alum, boric acid, sodium molybdate, sodium selenite, sodium tungstate, nickel chloride, and derivatives thereof

[0205] xiii) 1, 2, 5, 10, 50 or 100 chemical elements or heavy metals selected from the group consisting of: cadmium, lead, arsenic, mercury, cobalt, vanadium, nickel, lithium, antimony, and copper, and derivatives thereof

[0206] xiv) 10 -50 , 10 -10 , 10 -5 , 10 -1 , 1, 10, 10 3 or 10 5gram chemical elements or heavy metals selected from the group consisting of cadmium, lead, arsenic, mercury, cobalt, vanadium, nickel, lithium, antimony and copper and their derivatives,

[0207] xv)10 -50 , 10 -10 , 10 -5 , 10 -1 , 1, 10, 10 3 or 10 5 at least one CMR, toxic or cytotoxic compound selected from the group consisting of nitrilotriacetic acid, manganese sulfate, cobalt nitrate, zinc sulfate, copper sulfate, potassium aluminum sulfate, boric acid, sodium molybdate, sodium selenite, sodium tungstate, nickel chloride and any derivatives thereof, and / or

[0208] xvi)10 -50 , 10 -10 , 10 -5 , 0.01, 10 -1 , 1, 10, 10 3 or 10 5 g of peptone.

[0209] The present invention also relates to a method according to the invention, wherein the at least one compound in the pre-growth and / or growth medium is present in a concentration of C2 or in a concentration of C 总量 =C1+C2, where:

[0210] - C1 is the concentration of at least one compound in the pre-growth and / or growth medium that is not consumed by the nanoparticle-producing cells,

[0211] - C2 is the concentration of at least one compound in the pre-growth and / or growth medium consumed by the nanoparticle-producing cells,

[0212] as well as,

[0213] - preferably, C1 and / or C2 are measured, separated or differentiated by using a method capable of separating at least one compound consumed by the nanoparticle-producing cells from at least one compound not consumed by the nanoparticle-producing cells (e.g. centrifugation or tangential filtration), and

[0214] - Preferably, C1 and / or C2 are measured or taken into account at the beginning, during or at the end of the pre-growth and / or growth phase.

[0215] In one embodiment of the invention, a compound in the pre-growth and / or growth medium consumed by the nanoparticle producing cells is, preferably, a compound contained within the nanoparticle producing cells when the nanoparticle producing cells consume such compound.

[0216] In another embodiment of the present invention, a compound in the pre-growth and / or growth medium that is not consumed by the nanoparticle-producing cells is, preferably when the nanoparticle-producing cells do not consume this compound, a compound included extracellularly of the nanoparticles.

[0217] In one embodiment, a method is used to separate the consumed and unconsumed compounds. This method preferably separates the whole bacteria from the pre-growth and / or growth medium that does not contain such bacteria. This method can be centrifugation or filtration, preferably tangential filtration, or a method capable of separating the whole bacteria from the liquid medium, based on the separation between the low-size and / or low-weight elements included in the pre-growth and / or growth medium and the large-size and / or large-weight elements in the whole bacteria.

[0218] The present invention also relates to the method according to the present invention, wherein the concentration of at least one compound included in the fed-batch medium, preferably iron, iron source, carbon, carbon source, nitrogen, and / or nitrogen source, is greater than 10 -6 μM, 1 μM, and / or 10 6 μM.

[0219] The present invention also relates to the method according to the present invention, wherein the pre-growth, growth, and / or fed-batch growth medium does not include at least one compound at a concentration that affects the growth of the nanoparticle-producing cells and / or the production of nanoparticles, and / or wherein the pre-growth, growth, and / or fed-batch growth medium is substantially free of at least one compound, wherein the at least one compound is selected from the group consisting of: 1) Wolfe vitamins or a medium containing a total number of different components higher than half of the Wolfe vitamins, 2) a component of the Wolfe vitamins, 3) folic acid, 4) pyridoxine, 5) riboflavin, 6) biotin, 7) thiamine, 8) niacin, 9) pantothenic acid, 10) vitamin B 12, 11) aminobenzoic acid, 12) lipoic acid, 13) Wolff's minerals or a culture medium containing more than half of the total number of different components of Wolff's minerals, 14) nitrilotriacetic acid, 15) magnesium sulfate, 16) sodium chloride, 17) manganese sulfate, 18) ferrous sulfate heptahydrate, 19) cobalt nitrate, 20) calcium chloride, 21) zinc sulfate heptahydrate, 22) copper sulfate pentahydrate, 23) potassium alum dodecahydrate, 24) boric acid, 25) sodium molybdate, 26) sodium selenite, 27) sodium tungstate, 28) yeast extract or a culture medium containing more than half of the total number of different components of yeast extract, 29) yeast extract equivalent or a culture medium containing more than half of the total number of different components of yeast extract equivalent, 30) 1, 2 or 5 proteins derived from or contained in yeast extract, 31) 1, 2 or 5 nucleic acids derived from or contained in yeast extract, 32) 1, 2 or 5 peptides or functional peptides derived from or contained in yeast extract, 33) glutathione, 34) dextran, 35) mannan, 36) trehalose, 37) flavor nucleotides derived from or contained in yeast extract, 38) B vitamins, 39) biotin, 40) 1, 2 or 5 volatile aromatic compounds derived from or contained in yeast extract, 41) chromium, 42) cobalt, 43) strontium, 44) nickel chloride, 45) or a culture medium containing more than half of the total number of different components of mineral elixir, 46) MnSO4, 47) NaCl, 48) FeSO4, 49) CoSO4, 50) CaCl2, 51) ZnSO4, 52) CuSO4, 53) KAl(SO4)2, 54) H3BO3, 55) Na2MoO4, 56) NiCl2, 57) Na2SeO3, 58) peptone or a culture medium containing more than half of the total number of different components of peptone, 59) a component of peptone, 60) 1, 2 or 5 proteins derived from or contained in peptone, (61) sugars derived from or contained in peptone, (62) amino acids derived from or contained in peptone, (63) ash derived from or contained in peptone, (64) fibers derived from or contained in peptone, 65) a CMR reagent, 66) boric acid, 67) an amino acid, 68) alanine, 69) arginine, 70) asparagine, 71) aspartic acid, 72) cysteine, 73) glutamine, 74) glutamic acid, 75) glycine, 76) histidine, 77) isoleucine, 78) leucine, 79) lysine, 80) methionine, 81) phenylalanine, 82) proline, 83) serine, 84) threonine, 85) tryptophan, 86) tyrosine, 87) valine, 88) a cytotoxic or toxic compound, 89) manganese sulfate, 90) copper sulfate, 91) potassium alum, 92) boric acid, 93) sodium tungstate, 94) a heavy metal different from iron, 95) titanium, 96) vanadium, 97) manganese, 98) nickel, 99) copper, 100) zinc, 101) gallium, 102) germanium,103) Arsenic, 104) Zirconium, 105) Niobium, 106) Molybdenum, 107) Technetium, 108) Ruthenium, 109) Rhodium, 110) Palladium, 111) Silver, 112) Cadmium, 113) Indium, 114) Tin, 115) Tellurium, 116) Lutetium, 117) Hafnium, 118) Tantalum, 119) Tungsten, 120) Rhenium, 121) Osmium, 122) Iridium, 123) Platinum, 125) Gold, 126) Mercury, 127) Thallium, 128) Lead, 129) Bismuth, 130) Polonium, 131) Astatine, 132) Lanthanum, 133) Cerium, 134) Praseodymium, 135) Neodymium, 136) Promethium, 137) Samarium, 138) Europium, 139) Gadolinium, 140) Terbium, 141) Dysprosium, 142) Holmium, 143) Erbium, 144) Thulium, 145) Ytterbium, 146) Actinium, 147) Thorium (Th), 148) Protactinium, 149) Uranium, 150) Neptunium, 151) Plutonium, 152) Americium, 153) Curium, 154) Berkelium, 155) Californium, 156) Einsteinium, 157) Fermium, 158) Nobelium, 159) Radium, 160) Lawrencium, 161) Rutherfordium (Rf), 162) Dubnium (Db), 163) Seaborgium (Sg), 164) Bohrium (Bh), 165) Hassium (Hs), 166), (Mt, Meitnerium), 167) Darmstadtium (Ds), 168) Roentgenium (Rg), 169) Copernicium (Cn), 170) Elements 113 - 118, 171) Helium, 172) Lithium, 173) Beryllium, 174) Boron, 175) Fluorine, 176) Aluminum, 177) Silicon, 178) Argon, 179) Scandium, 180) Chromium, 181) Nickel, 182) Copper, 183) Selenium, 184) Bromine, 185) Krypton, 186) Rubidium, 187) Yttrium, 188) Tin, 189) Antimony, 190) Iodine, 191) Xenon, 192) Cesium, 193) Barium, 194) Lutetium, 195) Astatine, 196) Radon, 197) Francium (Fr), 198) Mendelevium (Md), 199) Mt, 200) Ununbium (Uub), 201) Ununtrium (Uut), 202) Ununquadium (Uuq), 203) Ununpentium (Uup), 204) Ununhexium (Uuh), 205) Ununseptium (Uus), 206) Ununoctium (Uuo), 207) Salts of compounds 1) to 206), and 208) their derivatives.

[0220] The present invention also relates to a method according to the present invention, wherein the concentration of the compound that affects the growth of the nanoparticle - producing cells and / or nanoparticle production is: in the pre - growth, growth, and / or fed - batch culture medium: i) greater than 1 pM, 1 μM, 1 mM, 10 -50 M, 10 -10 or 10 -5 M per liter of pre - growth, growth, and / or fed - batch culture medium, or ii) greater than 10 -50 、10 -10 、10 -5 or 10 -3 grams of the compound per liter of pre - growth, growth, and / or fed - batch culture medium.

[0221] In one embodiment of the present invention, the concentration of the compound that affects the nanoparticle - producing cells and / or nanoparticle production is: in the pre - growth, growth, and / or fed - batch culture medium: i) less than 10 50 、1、10 -5 、10 -6 or 10 - 9 M per liter of pre - growth, growth, and / or fed - batch culture medium, or ii) less than 10 10 、1、10 -10or 10 -20 g of the compound per liter of pre-growth, growth and / or fed-batch medium.

[0222] The present invention relates to a method according to the present invention, wherein the growth medium and / or the fed-batch medium is supplemented with a fed-batch medium, and:

[0223] i) the pH value of the fed-batch medium is lower than the pH value of the pre-growth and / or growth medium, preferably at least 10 -5 , 0.1, 0.5, 1, 2, 3 or 5 pH units, and / or

[0224] ii) the concentration of at least one chemical element preferably selected from the group consisting of a) a phosphorus source or a phosphate source, b) a potassium source, c) a magnesium source, d) an iron source, e) a vitamin source, f) a calcium source, g) KH2PO4, h) MgSO4, i) FeCl3, j) thiamine, k) CaCl2 and l) its derivatives is greater in the fed-batch medium than in the pre-growth and / or growth medium, preferably at least 10 -50 、10 -10 、0、1.1、5、10 or 10 3 times.

[0225] In one embodiment, the above conditions (i) and (ii) are verified by considering the pH and concentration of at least one compound:

[0226] i) for the fed-batch medium, before injecting the fed-batch medium into the pre-growth and / or growth medium, and / or

[0227] ii) for the pre-growth and / or growth medium, before, during or after injecting the fed-batch medium into the pre-growth and / or growth medium.

[0228] The present invention also relates to a method according to the present invention, wherein:

[0229] - the nanoparticle-producing cells are magnetotactic bacteria, and / or

[0230] - the nanoparticles are magnetosomes.

[0231] In another embodiment of the present invention, the nanoparticles are high-purity nanoparticle-producing cells.

[0232] In another embodiment of the present invention, the nanoparticles are high-purity nanoparticles, preferably nanoparticles based on high-purity iron oxide, wherein the high-purity nanoparticles refer to nanoparticles in which the mass or number of atoms preferably selected from the group consisting of cadmium, lead, arsenic, mercury, cobalt, vanadium, nickel, lithium, antimony, copper and their derivatives is less than 100, 99, 90, 50, 20, 10, 1 or 0.1%. Among them, the nanoparticles based on high-purity iron oxide refer to high-purity nanoparticles in which the atomic number or mass of iron and / or oxygen and / or iron oxide is preferably higher than 1, 50, 90, 93 or 99%.

[0233] The present invention also relates to a method according to the present invention, wherein the pre-growth and / or growth medium comprises a calcium source, a carbon source, a nitrogen source, a phosphate or phosphorus source, a sulfur source, an iron source, a vitamin source and a calcium source, and:

[0234] - The concentration of the carbon source in the pre-growth and / or growth medium is greater than the concentration of at least one compound selected from the group consisting of a phosphate or phosphorus source, a sulfur source, a vitamin source and a calcium source in the pre-growth and / or growth medium, preferably at least 0, 0.5, 1.1, 2, 4, 10 or 100 times, and / or - The concentration of the nitrogen source in the pre-growth and / or growth medium is greater than the concentration of at least one compound selected from the group consisting of a phosphate or phosphorus source, a sulfur source, a vitamin source and calcium in the pre-growth and / or growth medium, preferably at least 0, 0.5, 1.1, 2, 4, 10 or 100 times.

[0235] The present invention also relates to a method according to the present invention, further comprising the step of storing, amplifying, preparing or injecting a cell bank of nanoparticle-producing cells in the pre-growth and / or growth and / or fed-batch medium, wherein, preferably, the storage, amplification or preparation of the cell bank is carried out in a cell bank medium preferably containing at least 10 -50 、10 -10 、10 -1 、0, 1, 5, 10, 50, 70, 90 or 99% (preferably in terms of the number of compounds) of the same compounds as the pre-growth and / or growth and / or fed-batch medium, and preferably at least one compound different from the compounds in the pre-growth and / or growth and / or fed-batch medium, such as vitamins, minerals, chelating agents, sucrose and / or cryoprotectants: i) not in the pre-growth and / or growth and / or fed-batch medium (ii) having a lower concentration in the pre-growth and / or growth and / or fed-batch medium than in the cell bank medium.

[0236] In one embodiment of the present invention, the nanoparticle-producing cell bank is a master cell bank, a working cell bank or a research cell bank of nanoparticle-producing cells. In some cases, such cell banks include more than 1, 10, 10 3 、1010 or 10 20 nanoparticle-producing cells, preferably per milliliter or per liter of pre-growth and / or growth and / or fed-batch medium. In some other cases, such a cell bank comprises less than 10 100 、10 50 、10 20 、10 10 or 10 3 nanoparticle-producing cells.

[0237] In another embodiment of the present invention, the cell bank is a combination of at least one cell for initiating the amplification of nanoparticle-producing particles in a pre-growth and / or growth and / or fed-batch medium.

[0238] In one embodiment of the present invention, the cell bank is prepared under conditions that are the same as or similar to the pre-growth and / or growth phase.

[0239] In another embodiment of the present invention, the cell bank is prepared by bubbling or injecting a gas into a cell bank medium with a lower oxygen concentration, preferably with an oxygen concentration of less than 50, 10 or 1% relative to the volume of the cell bank medium.

[0240] The present invention relates to a method according to the present invention, further comprising a purification stage for obtaining nanoparticles based on high-purity iron oxide, preferably starting from the nanoparticles separated from the nanoparticle-producing cells obtained at the end of the growth stage, wherein the purification stage preferably comprises removing at least one impurity from the nanoparticles produced in the growth stage using at least one heating stage, wherein, preferably, the temperature of the nanoparticles according to the present invention produced in the growth stage or substantially produced in the growth stage is first raised to temperature T i , and then preferably maintained at T hi for a heating time t i ranging from 1 second to 20 years, wherein T i is preferably from 50 °C to 700 °C.

[0241] In one embodiment of the present invention, T i is greater than -273, -100, -50, 0, 1, 10, 20, 50, 100, 200, 500, 700 or 10 3 °C.

[0242] In one embodiment of the present invention, T i is less than 10 20 、10 10 、10 5 、10 3 、100, 50, 10, 0, -10 or -50 °C.

[0243] In yet another embodiment of the present invention, T i is between 10 and 10 5 , between 50 and 10 4 , between 100 and 10 3 , between 150 and 700, or between 200 and 500 °C.

[0244] In one embodiment of the present invention, t hi is greater than 10 -50 , 10 -10 , 10 -5 , 10 -1 , 1, 10, 10 2 , 10 3 , 10 5 , or 10 10 seconds.

[0245] In another embodiment of the present invention, t hi is lower than 10 100 , 10 50 , 10 10 , 10 5 , 10 3 , 10, 5, 2, or 1 second.

[0246] In yet another embodiment of the present invention, t hi is between 10 -5 seconds and 1 year, between 1 second and 20 years, between 1 second and 1 year, between 1 second and 1 month, between 1 second and 1 week, between 1 second and 1 day, or between 1 second and 1 hour.

[0247] In one embodiment of the present invention, preferably, the time to increase the temperature from an initial temperature, which preferably corresponds to the temperature when the nanoparticles are injected into the device for heating the nanoparticles, to the temperature T i is shorter than the time t hi , preferably at least 1.1, 5, 10, or 10 3 times.

[0248] In another embodiment of the present invention, the time to increase the temperature to T i is longer than the time t hi , preferably at least 1.1, 5, 10, or 10 3 times.

[0249] In one embodiment of the present invention, the iron oxide-based nanoparticles are nanoparticles comprising preferably more than 1%, 50%, 70%, 90%, or 99% iron oxide by mass of the iron oxide, preferably regardless of the percentage of the coating or excipient material.

[0250] The present invention also relates, preferably, to cells for producing nanoparticles obtained by the method according to the present invention, said cells for producing nanoparticles having a purity of greater than 10 -10 、1、5、10、50、75、80、90、95、99 or 99.9% of:

[0251] i) iron, based on the ratio M FeC / M MC , where M FeC is the mass of iron in the nanoparticles based on high-purity iron oxide, and M MC is the mass of iron and metals or metalloids other than iron in the nanoparticles based on pure iron oxide,

[0252] ii) iron and at least one other metal selected from the group consisting of: sodium, magnesium, aluminum, potassium, calcium, scandium, titanium, chromium, manganese, zinc, gallium, strontium, yttrium, zirconium, niobium, molybdenum, technetium, indium, cesium, barium, lanthanum, cerium, praseodymium, neodymium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, lutetium, hafnium, rhenium and tungsten, based on the ratio M1 / M2, where M1 is the mass of iron and at least one other metal selected from the above group in the nanoparticles based on high-purity iron oxide, and M2 is the mass of all metals contained in the nanoparticles based on high-purity iron oxide, and / or

[0253] iii) iron and at least one other non-metal selected from the group consisting of: hydrogen, carbon, nitrogen, phosphorus, sulfur, fluorine, chlorine, bromine, iodine, helium, neon, argon, krypton, xenon, radon and oxygen, based on the ratio M3 / M4, where M3 is the mass of iron and at least one other non-metal selected from the above group in the nanoparticles based on high-purity iron oxide, and M4 is the mass of all chemical elements contained in the nanoparticles based on high-purity iron oxide.

[0254] The present invention also relates to cells for producing nanoparticles or nanoparticles based on high-purity iron oxide obtained by amplifying or growing cells for producing nanoparticles in a pure culture medium, said pure culture medium comprising less than 1, 10 -3 、10 -6 or 10 -9 % of at least one heavy metal selected from the group consisting of: cobalt, manganese, zinc, nickel, silver, aluminum, arsenic, barium, cadmium, chromium, copper, molybdenum (molybdate), lead, antimony, selenium, silicon (silica), titanium, thallium, mercury, vanadium, gold, iridium, osmium, rhodium, ruthenium, platinum, lithium, antimony, tin, tungsten and their derivatives, where the percentage is based on the ratio C FeM / C MM , where C FeM is the concentration of iron in the pure culture medium, and C MM is the concentration of iron and metals or metalloids other than iron in the pure culture medium.

[0255] The present invention also relates, preferably, to nanoparticles based on high-purity iron oxide obtained by the method of the present invention, said nanoparticles based on high-purity iron oxide comprising more than 10 -10 、1、5、10、50、75、80、90、93、95、99 or 99.9% of:

[0256] i) iron, based on the ratio of M FeN / M MN where M FeN is the mass of iron in the nanoparticles based on high-purity iron oxide, and M MN is the mass of iron and metals or metalloids other than iron in the nanoparticles based on high-purity iron oxide,

[0257] ii) iron and at least one other metal other than iron selected from the group consisting of: sodium, magnesium, aluminum, potassium, calcium, scandium, titanium, chromium, manganese, zinc, gallium, strontium, yttrium, zirconium, niobium, molybdenum, technetium, indium, cesium, barium, lanthanum, cerium, praseodymium, neodymium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, lutetium, hafnium, rhenium and tungsten, based on the ratio M1 / M2, where M1 is the mass of iron and at least one other metal selected from the above group in the high-purity iron oxide nanoparticles, and M2 is the mass of all metals contained in the high-purity iron oxide nanoparticles, and / or

[0258] iii) iron and at least one other non-metal selected from the group consisting of: hydrogen, carbon, nitrogen, phosphorus, sulfur, fluorine, chlorine, bromine, iodine, helium, neon, argon, krypton, xenon, radon and oxygen, based on the ratio M3 / M4, where M3 is the mass of iron and at least one other non-metal selected from the above group in the high-purity iron oxide nanoparticles, and M4 is the mass of all non-metals contained in the high-purity iron oxide nanoparticles.

[0259] The present invention also relates to producing cells according to the high-purity nanoparticles of the present invention, and / or producing cells according to the nanoparticles based on high-purity iron oxide of the present invention, wherein the metals or metalloids other than iron in the high-purity iron oxide nanoparticles and / or the high-purity nanoparticles for producing cells are selected from the group consisting of: sodium, magnesium, aluminum, potassium, calcium, scandium, titanium, chromium, manganese, zinc, gallium, strontium, yttrium, zirconium, niobium, molybdenum, technetium, indium, cesium, barium, lanthanum, cerium, praseodymium, neodymium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, lutetium, hafnium, rhenium and tungsten and their derivatives, at least 5 different metals or metalloids.

[0260] The present invention also relates to the nanoparticles based on high-purity iron oxide according to the present invention, wherein the nanoparticles based on high-purity iron oxide are magnetosomes.

[0261] The present invention also relates to a composition comprising the nanoparticles based on high-purity iron oxide according to the present invention.

[0262] The present invention also relates to cells for producing high-purity nanoparticles according to the present invention, also known as high-purity nanoparticle-producing cells, wherein the high-purity nanoparticle-producing cells are magnetotactic bacteria.

[0263] The present invention also relates to a composition comprising the high-purity nanoparticle-producing cells according to the present invention.

[0264] The present invention also relates to a composition comprising high-purity nanoparticle-producing cells and high-purity iron oxide nanoparticles preferably obtained by the method according to the present invention, wherein:

[0265] The high-purity nanoparticle-producing cells comprise more than 0, 1, 10, 50, 70, 90, 95 or 99% of:

[0266] i) iron, based on the ratio of M FeC / M MC where M FeC is the mass of iron in the high-purity nanoparticle-producing cells, and M MC is the mass of iron and metals or metalloids other than iron in the high-purity nanoparticle-producing cells,

[0267] ii) iron and at least one other metal selected from the group consisting of: sodium, magnesium, aluminum, potassium, calcium, scandium, titanium, chromium, manganese, zinc, gallium, strontium, yttrium, zirconium, niobium, molybdenum, technetium, indium, cesium, barium, lanthanum, cerium, praseodymium, neodymium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, lutetium, hafnium, rhenium and tungsten, based on the ratio M1 / M2, where M1 is the mass of iron and at least one other metal selected from the above group in the high-purity iron oxide nanoparticles, and M2 is the mass of all metals contained in the high-purity iron oxide nanoparticles, and / or

[0268] iii) iron and at least one other non-metal selected from the group consisting of: hydrogen, carbon, nitrogen, phosphorus, sulfur, fluorine, chlorine, bromine, iodine, helium, neon, argon, krypton, xenon, radon and oxygen, based on the ratio M3 / M4, where M3 is the mass of iron and at least one other non-metal selected from the above group in the high-purity iron oxide nanoparticles, and M4 is the mass of all non-metal elements contained in the high-purity iron oxide nanoparticles,

[0269] and

[0270] Based on the high-purity iron oxide nanoparticles, it comprises more than 0, 1, 10, 50, 75, 93, 99 or 99.9% of:

[0271] i) iron, based on the ratio of M FeN / M MN where M FeN is the mass of iron in the high-purity iron oxide nanoparticles, and M MNis the mass of iron and metals or metalloids other than iron in the high-purity iron oxide nanoparticles,

[0272] ii) iron and at least one other metal selected from the group consisting of: sodium, magnesium, aluminum, potassium, calcium, scandium, titanium, chromium, manganese, zinc, gallium, strontium, yttrium, zirconium, niobium, molybdenum, technetium, indium, cesium, barium, lanthanum, cerium, praseodymium, neodymium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, lutetium, hafnium, rhenium, and tungsten, based on the ratio M1 / M2, where M1 is the mass of iron and at least one other metal selected from the above group in the high-purity iron oxide nanoparticles, and M2 is the mass of all metals contained in the high-purity iron oxide nanoparticles, and / or

[0273] iii) iron and at least one other non-metal selected from the group consisting of: hydrogen, carbon, nitrogen, phosphorus, sulfur, fluorine, chlorine, bromine, iodine, helium, neon, argon, krypton, xenon, radon, and oxygen, based on the ratio M3 / M4, where M3 is the mass of iron and at least one other non-metal selected from the above group in the high-purity iron oxide nanoparticles, and M4 is the mass of all non-metal elements contained in the high-purity iron oxide nanoparticles.

[0274] The present invention also relates to a composition comprising high-purity nanoparticle-producing cells and high-purity iron oxide nanoparticles, wherein:

[0275] The high-purity nanoparticle-producing cells contain more than 0, 10 -50 、10 -10 、10 -5 、10 -2 、1、5、10、25、50、75、90、95、99 or 99.9% of:

[0276] i) iron, based on the ratio of M FeC / M MC where M FeC is the mass of iron in the high-purity nanoparticle-producing cells, and M MC is the mass of iron and metals or metalloids other than iron in the high-purity nanoparticle-producing cells,

[0277] ii) iron and at least one other metal selected from the group consisting of: sodium, magnesium, aluminum, potassium, calcium, scandium, titanium, chromium, manganese, zinc, gallium, strontium, yttrium, zirconium, niobium, molybdenum, technetium, indium, cesium, barium, lanthanum, cerium, praseodymium, neodymium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, lutetium, hafnium, rhenium, and tungsten, based on the ratio M1 / M2, where M1 is the mass of iron and at least one other metal selected from the above group in the high-purity iron oxide nanoparticles, and M2 is the mass of all metals contained in the high-purity iron oxide nanoparticles, and / or

[0278] iii) iron and at least one other non-metal selected from the group consisting of: hydrogen, carbon, nitrogen, phosphorus, sulfur, fluorine, chlorine, bromine, iodine, helium, neon, argon, krypton, xenon, radon, and oxygen, based on the ratio M3 / M4, where M3 is the mass of iron and at least one other non-metal selected from the above group in the high-purity iron oxide nanoparticles, and M4 is the mass of all non-metal elements contained in the high-purity iron oxide nanoparticles,

[0279] and

[0280] based on the high-purity iron oxide nanoparticles containing more than 0, 10 -50 、10 -10 、10 -5 、10 -3 、10 -1 、0、1、5、10、25、50、75、93、95、99 or 99.9% of:

[0281] i) iron, based on the ratio of M FeN / M MN where M FeN is the mass of iron in the high-purity iron oxide nanoparticles, and M MN is the mass of iron and metals or metalloids other than iron in the high-purity iron oxide nanoparticles,

[0282] ii) iron and at least one other metal other than iron selected from the group consisting of: sodium, magnesium, aluminum, potassium, calcium, scandium, titanium, chromium, manganese, zinc, gallium, strontium, yttrium, zirconium, niobium, molybdenum, technetium, indium, cesium, barium, lanthanum, cerium, praseodymium, neodymium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, lutetium, hafnium, rhenium, and tungsten, based on the ratio M1 / M2, where M1 is the mass of iron and at least one other metal selected from the above group in the high-purity iron oxide nanoparticles, and M2 is the mass of all metals contained in the high-purity iron oxide nanoparticles, and / or

[0283] iii) iron and at least one other non-metal selected from the group consisting of: hydrogen, carbon, nitrogen, phosphorus, sulfur, fluorine, chlorine, bromine, iodine, helium, neon, argon, krypton, xenon, radon, and oxygen, based on the ratio M3 / M4, where M3 is the mass of iron and at least one other non-metal selected from the above group in the high-purity iron oxide nanoparticles, and M4 is the mass of all non-metal elements contained in the high-purity iron oxide nanoparticles,

[0284] where the high-purity nanoparticle-producing cells and / or the iron oxide-based nanoparticles are preferably obtained by culturing nanoparticle-producing cells in a growth medium that is substantially free of at least one metal or non-metal, or contains less than 10M, or 1M, or 10 -1 M, or 10 -3 M, or 10-6 M, or at least one metal or non-metal at 1 nanomole, wherein the at least one metal or non-metal is selected from the group consisting of: 1) cadmium, 2) lead, 3) arsenic, 4) mercury, 5) cobalt, 6) vanadium, 7) nickel, 8) lithium, 9) antimony, 10) copper, 11) V (Valadium), 12) molybdenum (molybdate), 13) selenium, 14) barium, 15) chromium, 16) strontium, 17) radioactive chemical elements, 18) beryllium, 19) rubidium, 20) ruthenium, 21) rhodium, 22) palladium, 23) promethium, 24) ytterbium, 25) tantalum, 26) osmium, 27) iridium, 28) bismuth, 29) polonium, 30) francium, 31) radium, 32) actinium, 33) thorium, 34) protactinium, 35) uranium, 36) neptunium, 37) plutonium, 38) americium, 39) curium, 40) berkelium, 41) californium, einsteinium, 42) fermium, 43) mendelevium, 44) nobelium, 45) lawrencium, 46) rutherfordium, 47) dubnium, 48) seaborgium, 49) bohrium, 50) hassium, 51) 52) darmstadtium, 53) roentgenium, 54) copernicium, 55) niobium, 56) flerovium (Fl), 57) moscovium, 58) livermorium (Lv), 59) astatine, 60) tennessine (Ts), 61) oganesson (Og, Ao), and 62) their derivatives.

[0285] In one embodiment of the present invention, high-purity nanoparticle-producing cells and / or high-purity iron oxide-based nanoparticles can be obtained by culturing and / or amplifying nanoparticle-producing cells in the following:

[0286] a) a pre-growth and / or growth medium, and / or

[0287] b) a medium that does not contain at least one metal or metalloid other than iron, or does not contain at least one metal or metalloid other than iron at a concentration that affects the growth of nanoparticle-producing cells, and the metal or metalloid other than iron is preferably selected from the group consisting of cobalt, manganese, zinc, nickel, silver, aluminum, arsenic, barium, cadmium, chromium, copper, molybdenum, lead, antimony, selenium, silicon, tantalum, thallium, mercury, vanadium, gold, iridium, osmium, rhodium, ruthenium, platinum, lithium, antimony, tin, tungsten, and their derivatives.

[0288] The present invention also relates to high-purity nanoparticle-producing cells according to the present invention, and / or preferably, high-purity iron oxide nanoparticles obtained from the high-purity nanoparticle-producing cells according to the present invention, wherein:

[0289] - The high-purity nanoparticle-producing cells are magnetotactic bacteria, and / or

[0290] - The high-purity iron oxide nanoparticles are magnetosomes.

[0291] The present invention also relates to a composition, a medical device, a drug, a preparation, a suspension, a cosmetic composition, a plant composition, a biological composition, a mineral composition, and / or a nanoparticle composition, which comprises nanoparticle-producing cells according to the present invention for producing high-purity nanoparticles and / or high-purity iron oxide nanoparticles according to the present invention.

[0292] The present invention also relates to, based on the M FeC / M MC ratio, preferably comprising nanoparticle-producing cells for producing high-purity nanoparticles containing more than 1%, 25%, 50%, 75%, 90% or 99% iron, where M FeC is the mass of iron in the nanoparticle-producing cells for producing high-purity nanoparticles, M MC is the mass of iron and metals and metalloids other than iron in the nanoparticle-producing cells for producing high-purity nanoparticles, and / or based on the M FeN / M MN ratio, preferably comprising iron oxide-based nanoparticles containing more than 1, 10, 50, 75, 93 or 99% iron, where M FeN is the mass of iron in the high-purity iron oxide nanoparticles, M MN is the mass of iron and metals or metalloids other than iron in the high-purity iron oxide nanoparticles, where the nanoparticle-producing cells for producing high-purity nanoparticles and / or the iron oxide-based nanoparticles are preferably obtained by culturing nanoparticle-producing cells in a growth medium that is substantially free of metals selected from the group consisting of cadmium, lead, arsenic, mercury, cobalt, vanadium, nickel, lithium, antimony and copper.

[0293] The term "substantially free" herein means substances that are not intentionally added to the medium but may be present in the medium as impurities.

[0294] In certain cases, M FeC / M MC and / or M FeN / M MN may be less than 100, 99, 95, 90, 50, 25, 10, 5 or 1%.

[0295] The present invention also relates to a method for preparing high-purity iron oxide nanoparticles. Preferably, the method according to the present invention uses nanoparticle-producing cells amplified in a pre-growth stage and a subsequent growth stage to produce high-purity iron oxide nanoparticles, where:

[0296] a) The pre-growth stage has at least one of the following characteristics:

[0297] a1) It includes at least one sub-stage i, in which the nanoparticle-producing cells are amplified in a volume V PGSi containing a pre-growth medium.

[0298] a2)V PGSi The change between the start and the end of sub - stage i is not higher than 1, 10, 50, 80, 90 or 99%;

[0299] a3)i is preferably comprised between 1 and 5, 1 and 10 or 1 and 1000;

[0300] a4)V PGSi+1 / V PGSi is between 1.001 and 1000 or between 1.1 and 50;

[0301] a5) The duration of each sub - stage i is from 1 second to 1 year, 1 minute to 1 month, 10 minutes to 1 week or 60 minutes to 3 days;

[0302] a6) The duration of at least one sub - stage i is higher than 1 second or 1 hour;

[0303] a7) The temperature of the pre - growth medium in at least one sub - stage is 10 - 60 °C, 20 - 50 °C or 30 - 40 °C;

[0304] a8) The concentration of oxygen or air or compressed air in the pre - growth medium:

[0305] - From: a8i) at the start of at least one sub - stage i, the partial pressure of oxygen, air or compressed air, preferably greater than 50 mbar, most preferably equal to 210 mbar, or a8ii) at the start of at least one sub - stage i, the volume percentage of oxygen, air or compressed air, preferably equal to 25% to 100% relative to the maximum volume occupied by oxygen, air or compressed air in the saturated pre - growth medium, or a8iii) at the start of at least one sub - stage i, the volume of oxygen or air or compressed air, preferably 5 to 25% relative to the volume of the pre - growth medium,

[0306] - To: a8iv) at the end of at least one sub - stage i, the partial pressure of oxygen, air or compressed air, preferably less than 100 mbar, most preferably equal to 0 mbar, or a8v) at the end of at least one sub - stage i, the volume percentage of oxygen, air or compressed air, preferably equal to 0% to 50% relative to the maximum volume occupied by oxygen, air or compressed air in the saturated pre - growth medium, or a8vi) at the end of at least one sub - stage i, the volume of oxygen or air or compressed air, preferably equal to 0 to 10% relative to the volume of the pre - growth medium;

[0307] a9) During at least one sub - stage i of the pre - growth phase, preferably during at least 0, 1, 5, 10 or 50% of the entire duration of at least one sub - stage i, volume V is introduced PGSiThe amount of oxygen, air, or compressed air is less than 100 liters, 1000 milliliters, or 100 milliliters of oxygen, air, or compressed air per minute;

[0308] a10) During all or part of sub-stage i, the pre-growth medium is stirred at a speed of 0 to 100 revolutions per minute or 0 to 10 3 meters per minute, where the speed is preferably the speed of at least one compound in the pre-growth medium, whether in liquid, gaseous, or solid form;

[0309] a11) The pH value of the pre-growth medium is not maintained at a fixed pH value, preferably by not adding a fed-batch medium preferably containing an iron source or another medium preferably containing an iron source in addition to the pre-growth medium to the pre-growth medium;

[0310] a12) Between the start and end of at least one sub-stage i, the change in the pH value of the pre-growth medium is greater than 10 -5 、10 -1 、0.5 or 1 pH unit, preferably from a minimum value preferably less than 7 to a maximum value preferably greater than 7;

[0311] a13) By adding a fed-batch medium containing less than 10 5 、10 3 、10 2 、10 or 2 μM of iron or an iron source to the pre-growth medium, the pH of the pre-growth medium is maintained at 3 to 11 or 6 to 8;

[0312] a14) Between the start and end of at least one sub-stage i, the change in the total concentration of at least one of iron, an iron source, carbon, a carbon source, nitrogen, and a nitrogen source in the pre-growth medium is less than 100, 50, 20, 10, 5, or 1%;

[0313] a15) The total concentration of iron or an iron source contained in the pre-growth medium is less than 10 5 mM, 10 3 mM, 10 mM, 2 mM of iron or an iron source per liter of pre-growth medium or 10 5 、10 3 、10 2 、50, 10, 5, 2, 1, 0.5 g of iron or an iron source per liter of pre-growth medium;

[0314] a16) The total concentration of iron or an iron source contained in the pre-growth medium is greater than 10 -50 M or 1 pM of iron or an iron source per liter of pre-growth medium or 0.4 ng of iron or an iron source per liter of pre-growth medium;

[0315] a17) The total concentration of carbon or carbon source contained in the pre-growth medium is less than 2 M or 260 g of carbon or carbon source per liter of pre-growth medium;

[0316] a18) The total concentration of carbon or carbon source contained in the pre-growth medium is greater than 0.1 nM or 0.1 ng of carbon or carbon source per liter of pre-growth medium;

[0317] a19) The total concentration of nitrogen or nitrogen source contained in the pre-growth medium is less than 740 mM or 40 g of nitrogen or nitrogen source per liter of pre-growth medium;

[0318] a20) The total concentration of nitrogen or nitrogen source contained in the pre-growth medium is greater than 0.1 nM or 0.1 ng of nitrogen or nitrogen source per liter of pre-growth medium;

[0319] a21) Between the start and end of at least one sub-stage i, the amount or concentration or percentage of carbon or carbon source consumed by the nanoparticle-producing cells is greater than:

[0320] ·10 -50 、0.01, 1 or 10 g of carbon or carbon source per liter of pre-growth medium, or 1 mM of carbon or carbon source per liter of pre-growth medium, and / or

[0321] ·10 -10 、1, 50 or 75%, where the percentage is preferably based on the ratio (Q Cf -Q Ci ) / Q Ci where Q Cf and Q Ci are respectively the amounts of carbon contained in the pre-growth medium at the end and start of at least one sub-stage i,

[0322] a22) Between the start and end of at least one sub-stage i, the amount or concentration or percentage of nitrogen or nitrogen source consumed by the nanoparticle-producing cells is greater than:

[0323] ·10 -50 、0.001, 1 or 10 g of nitrogen or nitrogen source per liter of pre-growth medium, or 0.1 mM of nitrogen or nitrogen source per liter of pre-growth medium, and / or

[0324] ·10 -10 、1, 50 or 75%, where the percentage is preferably based on the ratio (Q Nf -Q Ni ) / Q Ni where Q Nf and Q Ni are respectively the amounts of nitrogen contained in the pre-growth medium at the end and start of at least one sub-stage i,

[0325] a23) Between the start and end of at least one sub-stage i, the amount, concentration, or percentage of iron or iron source consumed by the nanoparticle-producing cells is greater than:

[0326] ·10 -10 、0.0001, 1, 10 or 10 10 mg of iron or iron source per liter of pre-growth medium, or 0.5 μM of iron or iron source per liter of pre-growth medium, and / or

[0327] ·10 -10 、1, 20, 50 or 75%, where the percentage is preferably based on the ratio (Q Fef -Q Fei ) / Q Fei , where Q Fef and Q Fei are respectively the amounts of iron contained in the pre-growth medium at the end and start of at least one sub-stage i;

[0328] a24) The amounts of carbon, carbon source, nitrogen, nitrogen source, iron and / or iron source consumed by the nanoparticle-producing cells between the start and end of at least one sub-stage i are less than the total concentrations of carbon, carbon source, nitrogen, nitrogen source, iron and / or iron source in the pre-growth medium;

[0329] a25) Between the start and end of sub-stage i, the total concentration change of carbon, carbon source, nitrogen, nitrogen source, iron and / or iron source in the pre-growth medium is not higher than 1, 10, 20, 50, 80 or 99%;

[0330] a26) Between the start and end of sub-stage i, the concentration increase of carbon, carbon source, nitrogen, nitrogen source, iron and / or iron source consumed by the nanoparticle-producing cells is higher than 1, 10, 20, 50, 80, 90 or 99%;

[0331] where these conditions preferably result in the nanoparticle-producing cells having at least one of the following characteristics:

[0332] a27) The nanoparticle-producing cells produce substantially no nanoparticles, or the nanoparticle-producing cells produce less than 10 10 、10 5 、1, 10 -3 or 10 -6 mg of nanoparticles per liter of pre-growth medium, where the quantity is preferably the number of nanoparticles produced at the end of at least one sub-stage i, or the difference between the number of nanoparticles produced at the end of at least one sub-stage i and the number of nanoparticles produced at the start of at least one sub-stage i;

[0333] a28) Preferably without concentration at the end of at least one sub-stage i, preferably between 0 and 10 5nm measurement, most preferably at 565 nm, of the optical density produced by nanoparticle-producing cells, characterized by at least one of the following properties:

[0334] a28i) During at least one sub-phase i of the pre-growth phase, it varies within an optical density range between 10 -20 and 10 20 or between 0.0001 and 40;

[0335] a28ii) Between the start and end of at least one sub-phase i, it increases by a factor higher than 0, 0.5, 1, 1.1, 5, 10, 10 3 or 10 5 times, where the factor is preferably the ratio of the optical density measured at the end of sub-phase i to the optical density measured at the start of sub-phase i;

[0336] a28iii) Between the start and end of at least one sub-phase i, it increases by a factor less than 10 10 、10 5 、2000、10 3 、10 2 、10、5、2 or 1 times;

[0337] a28iv) At the end of at least one sub-phase i, it has a maximum value less than 10 5 、10 3 or 100;

[0338] a28v) At the start of at least one sub-phase i, it has a minimum value greater than 0, 10 -50 、10 -10 、0.0001、10 -3 or 10 -1 ;

[0339] a29) The nanoparticle-producing cells are characterized in that, preferably during at least one sub-phase i of the pre-growth phase, the doubling time or the duration for the quantity to be multiplied by 2 is:

[0340] a29i) Greater than 10 -50 、10 -5 、1、10、10 2 or 10 3 minutes;

[0341] a29ii) Less than 10 3 、10、1 or 0.1 months; and / or

[0342] a29iii) 1 second to 1 month or 1 minute to 1 month;

[0343] b) A growth phase, including amplifying nanoparticle-producing cells derived from and / or generated during the pre-growth phase in at least one growth phase, preferably only in one growth phase. Most preferably, the number of growth phases is less than the number of sub-phases i of the pre-growth phase, wherein at least one growth phase has at least one property selected from the group consisting of:

[0344] b1) The nanoparticle-producing cells are amplified in a volume V of a growth medium GS ;

[0345] b2) V GS is larger than the volume of at least one sub-phase i of the pre-growth phase, preferably at least 0, 1, 1.1, 5, 10 or 10 3 times;

[0346] b3) V GS equals: V GS0 + V FB , where V GS0 is the volume of the growth medium at the start of the growth phase, and V FB is the volume of the fed-batch medium added to the growth medium during the growth phase;

[0347] b4) V GS varies by more than 10 -3 , 10 -1 , 1, 5, 10, 25, 50 or 75% between the start and end of at least one growth phase, preferably by adding a fed-batch medium to the growth medium during the growth phase;

[0348] b5) The volume V of the growth medium at the start of at least one growth phase GS0 is greater than the volume V of the fed-batch medium added to the growth medium during at least one growth phase FB , preferably at least 0, 1, 1.1, 1.5, 2, 5, 10 or 10 3 times;

[0349] b6) The duration of at least one growth phase is between 1 minute and 1 month, preferably between 40 hours and 15 days,

[0350] b7) The duration of at least one growth phase is greater than the duration of at least one sub-phase i of the pre-growth phase, preferably at least 0, 1, 1.1, 2, 5, 10 or 10 3 times,

[0351] b8) The temperature of the growth medium is 10 - 60 °C, 20 - 50 °C or 30 - 40 °C;

[0352] b9) Concentration of oxygen, air or compressed air in the growth medium:

[0353] From: a9i) at the start of at least one growth stage, preferably a partial pressure of oxygen, air or compressed air greater than 1 or 10 mbar, most preferably equal to 210 mbar, or a9ii) at the start of at least one growth stage, a volume percentage of oxygen, air or compressed air preferably equal to 10% to 100% relative to the maximum volume occupied by oxygen, air or compressed air in the saturated growth medium, or a9iii) at the start of at least one growth stage, a volume of oxygen or air or compressed air preferably 1 to 25% relative to the volume of the growth medium,

[0354] - Decrease to: a9iv) at the end of at least one growth stage, preferably a partial pressure of oxygen, air or compressed air less than 50 or 500 mbar, most preferably equal to 0 mbar, or a9v) at the end of at least one growth stage, a volume percentage of oxygen, air or compressed air preferably equal to 0% to 25% relative to the maximum volume occupied by oxygen, air or compressed air in the saturated growth medium, or a9vi) at the end of at least one growth stage, a volume of oxygen or air or compressed air preferably equal to 0 to 5% relative to the volume of the growth medium.

[0355] b10) During the entire growth stage or part of the growth stage, preferably for more than 1% of the total duration of the growth stage, introduce an amount of oxygen, air or compressed air with a volume V GS greater than 1, 10 or 200 ml of oxygen, air or compressed air per minute;

[0356] b11) At the end of the growth stage, introduce an amount of oxygen, air or compressed air with a volume V GS that is greater than at the start of the growth stage, preferably at least 0, 1, 1.1, 5, 10 or 10 3 times;

[0357] b12) During the entire growth stage or part of the growth stage, introduce an amount of oxygen, air or compressed air with a volume V GS such that the amount increases, with an increase greater than 10 -10 per minute, 10 -5 per minute, 1, 10 or 10 5 mL of oxygen, air or compressed air;

[0358] b13) During the entire growth stage or part of the growth stage, preferably at (greater than the pre-growth medium) greater than 1, 10 or 100 revolutions per minute or 0 to 10 per minute 3Stir the growth medium at a speed of meters, where the speed is preferably the speed of at least one compound in the growth medium, whether in liquid, gaseous or solid state;

[0359] b14) The pH value of the growth medium changes less than that of the pre-growth medium, or is maintained at a fixed pH value between 1 and 14 or between 6.5 and 7.5, or the pH value change is prevented from being higher than 0.1, 0.5 or 10 pH units, preferably by adding a fed-batch medium or another medium other than the growth medium to the growth medium;

[0360] b15) Between the start and the end of the growth phase, the pH value of the growth medium changes less than 10, 0.5 or 0.1 pH units, preferably from a minimum value less than 7.5 to a maximum value preferably greater than 6.5.

[0361] b16) The total concentration of carbon, carbon source, nitrogen, nitrogen source, iron and / or iron source contained in the growth medium changes by more than 0, 10 -5 , 1 or 50% between the start and the end of at least one growth phase,

[0362] b17) The total concentration of carbon or carbon source contained in the growth medium is greater than 10 -5 mM or 0.1 mM or 10 -5 g or 0.01 g of carbon or carbon source per liter of growth medium;

[0363] b18) The total concentration of carbon or carbon source contained in the growth medium is lower than 10 3 M or 2 M or 10 3 g or 180 g of carbon or carbon source per liter of growth medium;

[0364] b19) The total concentration of nitrogen or nitrogen source contained in the growth medium is greater than 10 -50 mM or 0.01 mM or 5·10 -10 g or 0.00005 g of nitrogen or nitrogen source per liter of growth medium;

[0365] b20) The total concentration of nitrogen or nitrogen source contained in the growth medium is lower than 10 10 mM or 111 mM or 10 5 g or 6 g of nitrogen or nitrogen source per liter of growth medium;

[0366] b21) The total concentration of iron or iron source contained in the growth medium is greater than 10 -5 nM or 1 nM or 10 -10 g or 3·10 -7 g of iron or iron source per liter of growth medium;

[0367] b22) The total concentration of iron or iron source contained in the growth medium is lower than 10 5or 1 mM or 10 5 g or 0.3 g of iron or iron source per liter of growth medium;

[0368] b23) Between the start and the end of at least one growth stage, the amount, concentration or percentage of carbon or carbon source consumed by the nanoparticle-producing cells is greater than:

[0369] ·10 -10 、10 -5 、0.1、1、10 or 10 3 g of carbon or carbon source per liter of growth medium, or 1 mM of carbon or carbon or carbon source per liter of growth medium, and / or

[0370] ·10 -50 、10 -10 、10 -5 、10 -1 、1、5、10、50 or 75%, where the percentage is preferably based on the ratio (Q Cf -Q Ci ) / Q Ci , where Q Cf and Q Ci are the amounts of carbon contained in the growth medium at the end and the start of at least one growth stage, respectively,

[0371] b24) Between the start and the end of at least one growth stage, the amount, concentration or percentage of nitrogen or nitrogen source consumed by the nanoparticle-producing cells is greater than:

[0372] ·10 -10 、10 -5 、0.01、1 or 10 g of nitrogen or nitrogen source per liter of growth medium, or 0.6 mM of nitrogen or nitrogen source per liter of growth medium, and / or

[0373] ·10 -50 、10 -10 、10 -5 、10 -1 、1、5、10、50 or 75%, where the percentage is preferably based on the ratio (Q Nf -Q Ni ) / Q Ni , where Q Nf and Q Ni are the amounts of nitrogen contained in the growth medium at the end and the start of at least one growth stage, respectively,

[0374] b25) Between the start and the end of at least one growth stage, the amount, concentration or percentage of iron or iron source consumed by the nanoparticle-producing cells is greater than:

[0375] ·10 -10 、10 -5, 0.01 or 1 mg of iron or an iron source per liter of growth medium, or 0.04 μM of iron or an iron source per liter of growth medium, and / or

[0376] ·10 -50 , 10 -10 , 10 -5 , 10 -1 , 1, 5, 10, 50 or 75%, where the percentage is preferably based on the ratio (Q Fef -Q Fei ) / Q Fei , where Q Fef and Q Fei are respectively the amounts of iron contained in the growth medium at the end and the beginning of at least one growth stage;

[0377] b26) Between the beginning and the end of at least one growth stage, preferably the amount of iron or an iron source introduced into the growth medium through a fed-batch medium is greater than 10 -10 mg or 0.3 mg of iron or an iron source per liter of growth medium, or greater than 10 -3 μM or 1 μM of iron or an iron source per liter of growth medium;

[0378] b27) Between the beginning and the end of at least one growth stage, preferably the amount of carbon or a carbon source introduced into the growth medium through a fed-batch medium is greater than 10 -50 , 10 -10 , 10 -5 or 0.07 g of carbon or a carbon source per liter of growth medium, or greater than 10 -10 , 10 -5 , 10 -3 , 0.8, 1 or 10 3 mM of carbon or a carbon source per liter of growth medium;

[0379] b28) Between the beginning and the end of at least one growth stage, preferably the amount of nitrogen or a nitrogen source introduced into the growth medium through a fed-batch medium is greater than 10 -10 , 0.006 or 1 g of nitrogen or a nitrogen source per liter of growth medium or 0.4 mM of nitrogen or a nitrogen source per liter of growth medium;

[0380] b29) Between the beginning and the end of at least one growth stage, the amounts of carbon, carbon source, nitrogen, nitrogen source, iron and / or iron source consumed by the nanoparticle-producing cells are less than the total concentrations of carbon, carbon source, nitrogen, nitrogen source, iron and / or iron source in the growth medium;

[0381] where these conditions preferably result in the nanoparticle-producing cells having at least one of the following characteristics:

[0382] b30) The nanoparticle-producing cells produce nanoparticles, or the nanoparticles produced by the nanoparticle-producing cells are greater than 10-50 、 10 -10 、 0.01 or 1 mg per liter of growth medium, where this quantity is preferably the quantity of nanoparticles produced at the end of the growth phase, or the difference between the quantity of nanoparticles produced at the end of the growth phase and the quantity of nanoparticles produced at the start of the growth phase;

[0383] b31) Preferably without concentration at the end of at least one growth phase, preferably measured between 0 and 10 4 nm, most preferably at 565 nm, the optical density produced by the nanoparticle-producing cells, characterized by at least one of the following properties:

[0384] b31i) The optical density at the end of the growth phase is greater than the optical density at the end of at least one sub-phase i of the pregrowth phase, preferably at least 0, 0.5, 1, 1.1, 1.5, 2, 5, 10 or 10 3 times;

[0385] b31ii) The optical density of the growth phase varies within an optical density range between 0.001 and 300;

[0386] b31iii) Between the start and end of at least one growth phase, the optical density increases by a factor higher than 0, 0.5, 1, 1.1, 5, 10 or 10 3 times, where this factor is preferably the ratio between the optical density measured at the end of at least one growth phase and the optical density measured at the start of at least one growth phase;

[0387] b31iv) Between the start and end of at least one growth phase, the optical density increases by a factor less than 10 10 、 10 4 or 10 times;

[0388] b31v) At the end of at least one growth phase, the optical density has a maximum value less than 10 10 、 10 5 、 300 or 10;

[0389] b31vi) At the start of at least one growth phase, the optical density has a minimum value greater than 10 -50 、 10 -10 、 0.001 or 0.01; and / or

[0390] b32) The nanoparticle-producing cells are characterized in that, preferably during the entire growth phase or part of the entire growth phase, the doubling time or the duration of multiplying the value by 2 is:

[0391] b32i) Greater than 1 minute;

[0392] b32ii) less than 1 month;

[0393] b32iii) from 1 minute to 1 month; and / or

[0394] b32iv) lower than the doubling time of at least one sub-stage i of the pre-growth stage, preferably by a factor of at least 1.1;

[0395] wherein, preferably, preferably measured per liter of pre-growth, growth and / or fed-batch medium, the pre-growth, growth and / or fed-batch medium does not contain more than: i) 1, 2, 3 or 6 different vitamins, ii) 10 -9 mol of at least one vitamin, iii) 10 -4 g of yeast extract, iv) 10 -9 mol of a component of at least one yeast extract, v) 1, 2, 5 or 10 components of yeast extract, vi) 10 -5 g of peptone, vii) 1 or 2 different CMR reagents, viii) 0.05 mg of at least one CMR reagent, ix) 1, 2 or 5 different chelating agents, x) 10 -8 mol of at least one chelating agent, xi) 1, 2 or 5 different amino acids, xii) 1 mg of at least one amino acid, xiii) 1, 2 or 5 different toxic or cytotoxic compounds, xiv) 1 mg of at least one toxic or cytotoxic compound, xv) 1, 3 or 7 different heavy metals other than iron, xvi) 1 mg of at least one heavy metal other than iron, xvii) more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 metals or chemical elements selected from cadmium, lead, arsenic, mercury, cobalt, vanadium, nickel, lithium, antimony and copper, xviii) 1 mg of cadmium, lead, arsenic, mercury, cobalt, vanadium, nickel, lithium, antimony and / or copper, xix) 0.5 mL or 10 -8 mol of Wolfe vitamins, xx) 10 -9 mol of a component of at least one Wolfe vitamin, xxi) 1, 2, 5 or 10 different components of Wolfe vitamins, xxii) 5 ml or 10 -8 mol of Wolfe minerals, xxiii) 10 -8 mol of a component of at least one Wolfe mineral, xxiv) 1, 2, 5, 7, 10 or 15 different components of Wolfe minerals, xxv) 10 -9 mol of mineral elixir, xxvi) 10 - 9 mol of a component of at least one mineral elixir, and / or xxvii) 1, 2, 5, 10 or 14 different components of mineral elixirs.

[0396] The invention also relates to a method according to the invention, wherein the end of the pre-growth and / or growth phase is characterized by at least one of the following properties:

[0397] i) The optical density of the pre-growth and / or growth medium saturates, or increases by less than 1 O.D. unit per hour during the pre-growth and / or growth phase;

[0398] ii) The biomass of the pre-growth and / or growth medium saturates, or increases by less than 1 g of nanoparticle-producing cells per hour during the pre-growth and / or growth phase;

[0399] iii) The nanoparticle-producing cells stop producing nanoparticles, or produce less than 0.01 mg of nanoparticles per hour during the pre-growth and / or growth phase;

[0400] iv) The fed-batch medium is no longer added to the pre-growth and / or growth medium;

[0401] v) Oxygen is no longer bubbled into the pre-growth and / or growth medium;

[0402] The invention also relates to a method according to the invention, wherein the start of the pre-growth and / or growth phase is characterized by at least one of the following properties:

[0403] i) The optical density of the pre-growth and / or growth medium starts to increase, preferably by more than 0.1 O.D. unit per day during the pre-growth and / or growth phase;

[0404] ii) The biomass of the pre-growth and / or growth medium starts to increase, preferably by more than 0.1 g of nanoparticle-producing cells per day during the pre-growth and / or growth phase;

[0405] iii) The nanoparticle-producing cells start to produce nanoparticles, preferably more than 0.01 mg of nanoparticles per day during the pre-growth and / or growth phase;

[0406] iv) The fed-batch medium starts to be added to the pre-growth and / or growth medium;

[0407] v) Oxygen starts to be bubbled into the pre-growth and / or growth medium.

[0408] The invention also relates to a method according to the invention, wherein the pre-growth, growth and / or fed-batch medium compositions are defined hereinafter.

[0409] Preferably, the pre-growth medium comprises at least 1, 2, 3, 4, 5, 6 or 7 of the following chemical elements or derivatives, and preferably the derivatives are or comprise a carbon source, a nitrogen source, a phosphate (phosphorus) source, a magnesium source, a potassium source, a calcium source, a vitamin source and / or a chlorine source: i) sodium lactate or a carbon source, preferably at a concentration of 10 -20or 0.0023 mol or g of sodium lactate or carbon source per liter of pre-growth medium and 0.23 or 10 5 mol or g of sodium lactate or carbon source per liter of pre-growth medium, with the most preferred concentration being 0.023 ± 0.01 mol of sodium lactate or carbon source per liter of pre-growth medium, ii) ammonium chloride or nitrogen source, preferably at a concentration between 10 -20 or 7.4·10 -4 mol or g of ammonium chloride or nitrogen source per liter of pre-growth medium and 7.4·10 -2 or 10 5 mol or g of ammonium chloride or nitrogen source per liter of pre-growth medium, with the most preferred concentration being (7.5 ± 1)·10 -3 mol of ammonium chloride or nitrogen source per liter of pre-growth medium, iii) KH2PO4 or potassium source or phosphorus source, preferably at a concentration between 10 -20 or 1.55·10 -5 mol or g of KH2PO4, potassium source or phosphorus source per liter of pre-growth medium and 1.55·10 -3 or 10 5 mol or g of KH2PO4 or potassium source or phosphorus source per liter of pre-growth medium, with the most preferred concentration being (1.5 ± 1)·10 - 4 mol of KH2PO4 or potassium source or phosphorus source per liter of pre-growth medium, iv) MgSO4 or magnesium source, preferably at a concentration between 10 -20 or 4.1·10 -5 mol or g of MgSO4 or magnesium source per liter of pre-growth medium and 4.1·10 -3 or 10 5 mol or g of MgSO4 or magnesium source per liter of pre-growth medium, with the most preferred concentration being (4 ± 1)·10 -4 mol of MgSO4 or magnesium source per liter of pre-growth medium, v) iron source or FeCl3, preferably at a concentration between 10 -20 or 2·10 -7 mol or g of FeCl3 or iron source per liter of pre-growth medium and 10 -5 or 10 5 mol or g of FeCl3 or iron source per liter of pre-growth medium, with the most preferred concentration being (2 ± 1)·10 -6 mol of FeCl3 or iron source per liter of pre-growth medium, vi) thiamine or vitamin, preferably at a concentration between 10 -20 or 8·10 -9 mol or g of thiamine or vitamin per liter of pre-growth medium and 8·10 -7 or 10 5 mol or g of thiamine or vitamin per liter of pre-growth medium, with the most preferred concentration being (8 ± 2)10-8 mol thiamine or vitamin per liter of pre - growth medium, vii) CaCl2 or calcium source or chlorine source, preferably at a concentration of 10 -20 or 10 -5 mol or g of CaCl2 or calcium source or chlorine source per liter of pre - growth medium and 10 -3 or 10 5 mol or g of CaCl2 or calcium source or chlorine source per liter of pre - growth medium, with the optimal concentration being (1 ± 0.8)·10 -4 mol of CaCl2 or calcium source or chlorine source per liter of pre - growth medium.

[0410] Preferably, preferably before adding the fed - batch medium to the growth medium, the growth medium comprises at least 1, 2, 3, 4, 5, 6 or 7 of the following chemical elements or derivatives, and the preferred derivatives are or contain a carbon source, a nitrogen source, a phosphorus source, a magnesium source, a potassium source, a calcium source, a vitamin source and / or a chlorine source: i) sodium lactate or carbon source, preferably at a concentration of 10 -20 or 0.0014 mol or g of sodium lactate or carbon source per liter of growth medium and 0.14 or 10 5 mol or g of sodium lactate or carbon source per liter of growth medium, with the most preferred concentration being (0.014 ± 0.01) mol of sodium lactate or carbon source per liter of growth medium, ii) ammonium chloride or nitrogen source, preferably at a concentration of 10 -20 or 4.1·10 -4 mol or g of ammonium chloride or nitrogen source per liter of growth medium and 4.1·10 -2 or 10 5 mol or g of ammonium chloride or nitrogen source per liter of growth medium, with the most preferred concentration being (4.1 ± 1)·10 -3 mol of ammonium chloride or nitrogen source per liter of growth medium, iii) KH2PO4 or potassium source or phosphorus source, preferably at a concentration of 10 -20 or 1.55·10 -5 mol or g of KH2PO4 or potassium source or phosphorus source per liter of growth medium and 1.55·10 -3 or 10 5 mol or g of KH2PO4 or potassium source or phosphorus source per liter of growth medium, with the most preferred concentration being (1.5 ± 1)·10 -4 mol of KH2PO4 per liter of growth medium, iv) MgSO4 or magnesium source, preferably at a concentration of 10 -20 or 4.1·10 - 5 mol or g of MgSO4 or magnesium source per liter of growth medium and 4.1·10 -3 or 10 5 mol or g of MgSO4 or magnesium source per liter of growth medium, with the most preferred concentration being (4 ± 1)·10-4 mol of MgSO4 per liter of growth medium, v) FeCl3 or an iron source, preferably at a concentration of 10 -20 or 10 -7 mol of FeCl3 or an iron source per liter of growth medium and 10 -5 or 10 5 mol of FeCl3 or an iron source per liter of growth medium, most preferably at a concentration of (2 ± 1)·10 -6 mol of FeCl3 or an iron source per liter of growth medium, vi) thiamine or a vitamin, preferably at a concentration of 10 -20 or 8·10 -9 mol or g of thiamine or a vitamin per liter of growth medium and 8·10 -7 or 10 5 mol or g of thiamine or a vitamin per liter of growth medium, most preferably at a concentration of (8 ± 2)·10 -8 mol of thiamine or a vitamin per liter of growth medium, vii) CaCl2 or a calcium source or a chlorine source, preferably at a concentration of 10 -20 or 10 -5 mol or g of CaCl2 or a calcium source or a chlorine source per liter of growth medium and 10 -3 or 10 5 mol or g of CaCl2 or a calcium source or a chlorine source per liter of growth medium, the optimal concentration being (1 ± 0.8)·10 -4 mol of CaCl2 or a calcium source or a chlorine source per liter of growth medium.

[0411] Preferably, preferably before adding it to the growth medium, the fed-batch medium comprises at least 1, 2, 3, 4, 5, 6 or 7 of the following chemical elements or derivatives, preferably the derivatives being or comprising a carbon source, a nitrogen source, a phosphorus source, a magnesium source, a potassium source, a calcium source, a vitamin source and / or a chlorine source: i) lactic acid or a carbon source, preferably at a concentration of 10 -20 or 10 -1 mol or g of lactic acid or a carbon source per liter of fed-batch medium and 10 or 10 5 mol or g of lactic acid or a carbon source per liter of fed-batch medium, most preferably at a concentration of (1 ± 0.5) mol of lactic acid or a carbon source per liter of fed-batch medium, ii) ammonia or a nitrogen source, preferably at a concentration of 10 -20 or 2.8·10 -2 mol or g of ammonia or a nitrogen source per liter of fed-batch medium and 2.8 or 10 5 mol or g of ammonia or a nitrogen source per liter of fed-batch medium, most preferably at a concentration of (2.8 ± 1)·10 -1 mol of ammonia or a nitrogen source per liter of fed-batch medium, iii) KH2PO4 or a potassium source or a phosphorus source, preferably at a concentration of 10 -20 or 1.7·10-3 mol or g of KH2PO4 or potassium source or phosphorus source per liter of fed-batch medium and 1.7·10 -1 or 10 5 mol or g of KH2PO4 or potassium source or phosphorus source per liter of fed-batch medium, with the most preferred concentration being (1.7±1)·10 - 2 mol of KH2PO4 per liter of fed-batch medium, iv) MgSO4 or magnesium source, preferably at a concentration between 10 -20 or 2·10 -4 mol or g of MgSO4 or magnesium source per liter of fed-batch medium and 2·10 -2 or 10 5 mol or g of MgSO4 or magnesium source per liter of fed-batch medium, with the most preferred concentration being (2±1)·10 -3 mol of MgSO4 or magnesium source per liter of fed-batch medium, v) FeCl3 or iron source, preferably at a concentration between 10 -20 or 10 -4 mol or g of FeCl3 or iron source per liter of fed-batch medium and 10 -1 or 10 5 mol or g of FeCl3 or iron source per liter of fed-batch medium, with the most preferred concentration being (7±4)·10 -3 mol of FeCl3 or iron source per liter of fed-batch medium, vi) thiamine or vitamin, preferably at a concentration between 10 -20 or 10 -8 mol or g of thiamine or vitamin per liter of fed-batch medium and 10 -4 or 10 5 mol or g of thiamine or vitamin per liter of fed-batch medium, with the most preferred concentration being (2±1.5)·10 -6 mol of thiamine or vitamin per liter of fed-batch medium, vii) CaCl2 or calcium source or chlorine source, preferably at a concentration between 10 -20 or 10 -5 mol or g of CaCl2 or calcium source or chlorine source per liter of fed-batch medium and 10 -2 or 10 5 mol or g of CaCl2 or calcium source or chlorine source per liter of fed-batch medium, with the optimal concentration being (1±0.8)·10 -3 mol of CaCl2 or calcium source or chlorine source per liter of fed-batch medium.

[0412] The present invention also relates to a method according to the present invention, wherein the pre-growth, growth, and / or fed-batch medium comprises at least one source selected from the group consisting of:

[0413] - A carbon source selected from the group consisting of: at least one compound containing at least one carbon atom, lactic acid, sodium lactate, acetic acid, ethanolate, glucose, pyruvate, succinate, carbon dioxide, glycerol, and combinations thereof, with a concentration preferably from 1 nM to 2 mol / L;

[0414] - An iron source selected from the group consisting of: at least one compound containing at least one iron atom, iron citrate, iron quinate, iron chloride, iron sulfate, FeCl3, and combinations thereof, with a concentration preferably from 1 nM to 2·10 -3 mol / L;

[0415] - A nitrogen source selected from the group consisting of: at least one compound containing at least one nitrogen atom, nitrate, nitrogen gas, ammonium, ammonia, ammonium salts, urea, amino acids, ammonia gas, and combinations thereof, with a concentration preferably from 1 nM to 4 mol / L;

[0416] - An oxygen source selected from the group consisting of: at least one compound containing at least one oxygen atom, oxygen, oxygen gas, air, or compressed air, preferably in gaseous form. In some cases, the oxygen source is bubbled or introduced into the growth medium at a gas rate preferably between 5 milliliters of gas per minute and 50,000 milliliters of gas per minute.

[0417] - A phosphate source consisting of at least one compound containing at least one phosphate, with a concentration preferably from 1 nM to 2·10 -1 mol / L;

[0418] - A potassium source consisting of at least one compound containing at least one potassium atom, with a concentration preferably from 1 nM to 2·10 -1 mol / L;

[0419] - A sulfate source or sulfur source consisting of at least one compound containing at least one sulfur atom or sulfate, with a concentration preferably from 1 nM to 4·10 -1 mol / L;

[0420] - A manganese source consisting of at least one compound containing at least one manganese atom, with a concentration preferably from 1 nM to 4·10 -1 mol / L;

[0421] - A vitamin source selected from the group consisting of: at least one compound containing at least one vitamin, biotin, calcium, pantothenate, folic acid, inositol, niacin, p-aminobenzoic acid, pyridoxine hydrochloride, riboflavin, thiamine, thiamine hydrochloride, and their derivatives and combinations thereof, with a concentration preferably from 1 nM to 10 -4 mol / L, and

[0422] - Calcium source, consisting of at least one compound containing at least one calcium atom, with a concentration preferably ranging from 1 nM to 10 - 1 mol / L.

[0423] In one embodiment of the present invention, the sources of carbon, nitrogen, potassium, phosphorus, magnesium, calcium, vitamins, iron, oxygen, and / or chlorine preferably include at least 1, 2, 5, 10, or 10 3 carbon, nitrogen, potassium, phosphorus, magnesium, calcium, vitamin, iron, oxygen, and / or chlorine atoms within the chemical formula or molecule or composition of these sources.

[0424] In another embodiment of the present invention, when a compound or quantity or element or property P1 is higher, longer, or greater than a compound or quantity or element or property P2, this indicates that P1 = α·P2, where α is preferably a number or integer greater than 1, or P1 = α + P2, where α is preferably a number or integer greater than 0.

[0425] In another embodiment of the present invention, when a compound or quantity or element or property P1 is lower, shorter, or smaller than a compound or quantity or element or property P2, this indicates that P1 = P2 / α, where α is preferably a number or integer greater than 1, or P1 = P2 - α, where α is preferably a number or integer greater than 0.

[0426] In one embodiment of the present invention, the CMR compound is a carcinogenic, mutagenic, and / or reproductive toxic compound. In certain cases, a carcinogenic compound is a compound that preferably causes, induces, or is suspected of causing or inducing cancer in a living organism or the human body. In certain cases, a mutagenic compound is a compound that preferably causes, induces, or is suspected of causing or inducing at least one mutation, alteration, or change in the number or size of genes, DNA, RNA, DNA strands, RNA strands, and / or nucleic acids in a living organism or the human body. In certain cases, a reproductive toxic compound refers to a compound that preferably causes, induces, or is suspected of causing or inducing toxicity, mutation, alteration, or change in reproductive organs, embryos, or fetuses in a living organism or the human body.

[0427] In one embodiment of the present invention, a toxic or cytotoxic compound is a compound that preferably causes, induces, or is suspected of causing or inducing toxicity, death, weight loss, organ damage, behavioral changes, changes in food or water consumption, necrosis, apoptosis, cellular internalization, and changes in cell number, shape, and / or geometric structure in an individual or a living organism.

[0428] In certain cases, the compound is at a concentration greater than 10 -6 、10 -3 、10 -1 、1、10、10 3 or 106 at a concentration of μM is CMR or cytotoxic or toxic.

[0429] In some other cases, the compound is CMR or cytotoxic or toxic at a concentration of less than 10 20 、10 6 、10 3 、10、1、10 -1 、10 -3 or 10 -6 μM.

[0430] The present invention relates to a method for generating high-purity iron oxide nanoparticles using cells that generate nanoparticles, comprising:

[0431] i) a pre-growth step, which includes amplifying the cells that generate nanoparticles in a pre-growth medium such that the cells that generate nanoparticles substantially do not generate nanoparticles, or

[0432] ii) a growth step, which includes amplifying the cells that generate nanoparticles derived from the pre-growth step in a growth medium such that the cells that generate nanoparticles generate nanoparticles.

[0433] The present invention relates to a method for generating high-purity iron oxide nanoparticles using cells that generate nanoparticles, comprising:

[0434] i) a pre-growth step, which includes amplifying the cells that generate nanoparticles in a pre-growth medium such that the cells that generate nanoparticles substantially do not generate nanoparticles, and / or

[0435] ii) a growth step, which includes amplifying the cells that generate nanoparticles derived from the pre-growth step in a growth medium such that the cells that generate nanoparticles generate nanoparticles.

[0436] In one embodiment of the present invention, the growth medium is supplemented with a fed-batch medium.

[0437] In one embodiment of the present invention, the growth medium supplemented with the fed-batch medium is the growth medium.

[0438] In one embodiment of the present invention, the growth medium has at least one property in common with the pre-growth and / or fed-batch medium.

[0439] In one embodiment of the present invention, the growth medium and / or the pre-growth medium comprises at least one source selected from the group consisting of: i) a carbon source or sodium or lactate, preferably sodium lactate, ii) an ammonium source, preferably ammonium chloride, iii) a magnesium source, preferably magnesium sulfate, iv) a potassium source, preferably potassium phosphate, v) a vitamin source, preferably thiamine, vi) a calcium source, preferably calcium chloride, and vii) an iron source, preferably iron chloride. Each source in the growth and / or pre-growth medium preferably has a concentration included between 10 -6 and 10 3 mM, 10 -3 and 100 mM, 0.01 and 10 mM, or 0.1 and 10 mM.

[0440] In one embodiment of the present invention, in the pre-growth medium and / or the growth medium, the concentration of the vitamin and / or calcium and / or iron source is at least 0, 0.5, 0.1, 1.1, 1.2, 1.5, 5, 10, 10 3 or 10 5 times smaller than the concentration of the sodium source and / or ammonium source and / or magnesium source and / or potassium source.

[0441] In one embodiment of the present invention, the fed-batch medium comprises at least one source selected from the group consisting of: i) a carbon source, preferably lactic acid, ii) ammonia, iii) a potassium source, preferably potassium phosphate, iv) a magnesium source, preferably magnesium sulfate, v) an iron source, preferably iron chloride, vi) a vitamin source, preferably thiamine, vii) a calcium source, preferably calcium chloride, and vi) an iron source, preferably iron chloride. Each source in the growth and / or pre-growth medium preferably has a concentration included between 0.001 and 100 mM, 0.01 and 10 mM or 0.1 and 10 mM.

[0442] In one embodiment of the present invention, in the fed-batch medium, the concentration of the vitamin and / or calcium source is at least 0, 0.5, 1, 1, 1, 1, 2, 1, 5, 5, 10, 10 3 or 10 5 times smaller than the concentration of the sodium source and / or ammonium source and / or magnesium source and / or potassium source and / or iron source.

[0443] In one embodiment of the present invention, the nanoparticles described in the present invention are or comprise more than 1, 2, 5, 10, 10 3 、10 5 、10 10 、10 20 、10 50 or 10 100 components of nanoparticles, nanoparticles per liter of growth medium or nanoparticles per nanoparticle-producing cell. In some cases, iron oxide represents or is more than 1, 10, 10 3 、10 5, 10 10 , 10 20 , 10 50 or 10 100 iron atoms and / or greater than 1, 10, 10 3 , 10 5 , 10 10 , 10 20 , 10 50 or 10 100 oxygen atoms in the component. In some other cases, the chemical elements and / or impurities contained in the nanoparticles are or represent more than 1, 10, 10 3 , 10 5 , 10 10 , 10 20 , 10 50 or 10 100 chemical elements, and / or impurities contained in the nanoparticles.

[0444] In another embodiment of the present invention, the nanoparticles according to the present invention are or contain less than 10 100 , 10 50 , 10 20 , 10 10 , 10 5 , 10 3 , 100, 50, 10, 5 or 2 nanoparticles in the component, nanoparticles per liter of growth medium or nanoparticles per nanoparticle-generated cell. In some cases, iron oxide represents or is less than 1, 10, 10 3 , 10 5 , 10 10 , 10 20 , 10 50 or 10 100 iron atoms and / or less than 1, 10, 10 3 , 10 5 , 10 10 , 10 20 , 10 50 or 10 100 oxygen atoms. In some other cases, the chemical elements and / or impurities contained in the nanoparticles are or represent less than 10 100 , 10 50 , 10 20 , 10 10 , 10 5 , 10 3 , 100, 50, 10, 5 or 2 chemical elements and / or impurities contained in the nanoparticles.

[0445] In one embodiment of the present invention, the nanoparticles contain at least one impurity.

[0446] In one embodiment of the present invention, the high-purity iron oxide nanoparticles contain a small amount of impurities, for example when the method can obtain nanoparticles with a small amount of impurities. In some cases, the nanoparticles contain / do not contain at least one impurity or contain or contain or contain less than 10 50 、10 20 、10 10 、10 5 、10 2 、10, 5, 2, 5, 1, 10 -2 、10 -10 、10 -20 or 10 -50 impurities or impurities per gram of nanoparticles, or grams of impurities per gram of nanoparticles. In some other cases, the percentage of impurities (preferably by mass) contained inside or on the surface of the nanoparticles is less than 100, 90, 80, 70, 60, 50, 30, 20, 10, 5, 1, 0.1 or 0.001%. According to the present invention, in some cases, the impurity percentage can be defined as the ratio between the number of atoms, quantity, mass or volume of impurities contained in the nanoparticles divided by the total number of atoms, quantity, mass or volume of all chemical elements contained in the nanoparticles.. In some cases, all the chemical elements contained in the nanoparticles can be the sum of iron oxide, doping materials and impurities contained in the nanoparticles. In other cases, the impurity concentration contained inside or on the surface of the nanoparticles is less than 10 50 、10 30 、10 10 、10 5 、10 3 、500, 100, 50, 10, 1, 10 -1 、10 -3 、10 -5 、10 -10 or 10 -50 micrograms of impurities per gram of nanoparticles.

[0447] In another embodiment of the present invention, the high-purity iron oxide nanoparticles contain a large amount of impurities, for example when impurities are added or incorporated into the nanoparticles after preparing the nanoparticles by the method. In some cases, the nanoparticles contain more than 10 -50 、10 -20 、10 -10 、10 -5 、10 -2 、1, 2, 5, 10, 10 3 、10 5 、10 10 、10 20 or 10 50Impurities or grams of impurities per gram of nanoparticles or per gram of nanoparticles. In some cases, the nanoparticles contain a large amount of impurities. In some cases, the percentage (preferably by mass) of impurities contained inside or on the surface of the nanoparticles is greater than 10 -40 、10 -20 、10 -10 、10 -5 、10 -2 、10 -1 、1、5、10、25、50、75、80 or 90%. In some other cases, the concentration of impurities contained inside or on the surface of the nanoparticles is greater than 10 -100 、10 -50 、10 -20 、10 -10 、10 -5 、10 -3 、10 -2 、10 -1 、1、10、50、100、10 3 、10 5 or 10 10 micrograms of impurities per gram of nanoparticles.

[0448] In some cases, the impurities can be the same impurities, i.e., preferably impurities containing the same chemical element.

[0449] In some other cases, the impurities can be different impurities, i.e., preferably impurities containing at least one different chemical element.

[0450] In one embodiment of the present invention, the chemical elements are selected from: actinides, actinium, aluminum, americium, antimony, argon, arsenic, astatine, barium, berkelium, beryllium, bismuth, bismuth, boron, bromine, cesium, calcium, californium, carbon, cerium, chlorine, chromium, cobalt, cobalt, cadmium, copper, curium,

[0451] darmstadtium, darmstadtium, dysprosium, einsteinium, erbium, europium, cadmium, flerovium, fluorine, francium, gadolinium, gallium, germanium, gold, hafnium, helium, hassium, holmium, hydrogen, indium, iodine, iridium, iron, krypton, lanthanides, lanthanum, lawrencium, lead, lithium, livermorium, lutetium, magnesium, manganese, deuterium, mendelevium, mercury, molybdenum, neodymium, neon, neptunium, nickel, niobium, nitrogen, niobium, osmium, oxygen, palladium, phosphorus, platinum, plutonium, polonium, potassium, praseodymium, protactinium, promethium, radium, radon, rhenium, rhodium, roentgenium, rubidium, ruthenium, rutherfordium, samarium, selenium, silicon, silver, sodium, strontium, sulfur, scandium, selenium, tellurium, terbium, thorium, thulium, tin,

[0452] tantalum, technetium, thallium, titanium, tungsten, Og, moscovium, Ts (Tennessine), nihonium, uranium, vanadium, xenon, ytterbium, yttrium, zinc, zirconium, and combinations of several of these chemical elements.

[0453] The present invention also relates to a method according to the present invention, wherein the impurity is at least one chemical element different from iron, oxygen and / or iron oxide.

[0454] The present invention also relates to a method according to the present invention, wherein the impurity is preferably carbon or a carbonaceous material.

[0455] In one embodiment of the present invention, the carbonaceous material comprises at least one carbon atom, preferably but not necessarily mixed or assembled with other chemical elements other than carbon.

[0456] In yet another embodiment of the present invention, the carbon or carbonaceous material is derived from, produced by, or comes from cells that produce nanoparticles.

[0457] The present invention also relates to a method according to the present invention, wherein the nanoparticles obtained by the method comprise iron oxide, and the iron oxide has at least one of the following properties: i) it comprises at least one iron atom and one oxygen atom, ii) it forms a crystalline or mineral structure, iii) it may have the chemical formula FeO, FeO2, Fe3O4, Fe4O5, Fe5O6, Fe5O7, Fe 25 O 32 、Fe 13 O 19 、α-Fe2O3, β-Fe2O3, γ-Fe2O3, ε-Fe2O3, iv), it may be composed of wüstite, iron dioxide, magnetite, hematite, maghemite, v) it may be in the ε-phase, α-phase, β-phase, γ-phase, vi), it may be at various oxidation levels, vii) it has the formula Fe α O β D γ , where α, β and / or γ are coefficients, preferably stoichiometric coefficients. In some cases, α, β and / or γ are equal to 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 19 or 20. In some other cases, α, β and / or γ are greater than 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 19 or 20. In some other cases, α, β and / or γ are less than 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 19 or 20. In some other cases, D is a doping material of the nanoparticles. In some cases, the doping material may be selected from: aluminum, antimonite, barium, chromium, copper, gold, manganese, silver, tin, titanium and zinc.

[0458] In one embodiment of the present invention, the iron oxide contained in the nanoparticles is the main chemical element of the nanoparticles. In some cases, high-purity iron oxide nanoparticles may contain a large amount of iron oxide. In some cases, the percentage (preferably by mass) of iron oxide contained in the nanoparticles is greater than 10 -40 、10 -20 、10-10 , 10 -5 , 10 -2 , 10 -1 , 1, 5, 10, 25, 50, 75, 80, 90, 99 or 99.9%. According to the present invention, in some cases, the iron oxide percentage can be defined as the number of atoms, quantity, mass or volume of iron oxide in the iron oxide nanoparticles divided by the number of atoms, quantity, mass or volume of all chemical elements contained in the nanoparticles. In some other cases, the concentration of iron oxide contained in the nanoparticles is greater than 10 -100 , 10 -50 , 10 -20 , 10 -10 , 10 -5 , 10 -3 , 10 -2 , 10 -1 , 1, 10, 50, 100, 10 3 , 10 5 or 10 10 micrograms of iron oxide per gram of nanoparticles.

[0459] In one embodiment of the present invention, the high-purity iron oxide nanoparticles contain a small amount of iron oxide, for example, when the nanoparticles are treated and / or partially or completely destroyed and / or applied to an organism, or when a large amount of iron oxide is incorporated into the nanoparticles when the method cannot be achieved. In some cases, the percentage (preferably by mass) of iron oxide contained inside or on the surface of the nanoparticles is less than 100%, 90%, 80%, 70%, 50%, 30%, 10%, 5%, 0.1% or 0.001%. In some other cases, the concentration of iron oxide contained in the nanoparticles can be less than 10 50 , 10 30 , 10 10 , 10 5 , 10 3 , 500, 100, 50, 10, 1, 10 -1 , 10 -3 , 10 -5 , 10 -10 or 10 -50 micrograms of iron oxide per gram of nanoparticles.

[0460] In another embodiment of the present invention, the percentage, concentration, number of atoms, quantity, mass or volume ratio of iron oxide contained in the nanoparticles is greater than the percentage, concentration, number of atoms, quantity, mass or volume of impurities contained in the nanoparticles, preferably 1.00001, 1.001, 1.1, 2, 5, 10, 50, 10 2 , 10 3 , 10 5 , 1010 , 10 20 or 10 50 times.

[0461] In one embodiment of the present invention, the iron oxide and / or impurities are contained or incorporated in: i) within the nanoparticles, ii) on the surface of the nanoparticles, iii) outside the nanoparticles, iv) in the crystalline or amorphous structure of the nanoparticles, v) in the defects of the nanoparticles, and / or, vi) in the vacancies of the nanoparticles.

[0462] In one embodiment of the present invention, the iron oxide and / or impurities interact with the nanoparticles through, for example, electrostatic, strong, weak, nuclear, metallic, van der Waals, Debye, London or hydrogen bonds.

[0463] In one embodiment of the present invention, the iron oxide and / or impurities are located at a certain distance from the nanoparticles, preferably from the center or surface of the nanoparticles, less than 10 50 , 10 20 , 10 10 , 10 5 , 10 3 , 100, 10, 5 or 1 nm. In some cases, the center of the nanoparticle is a region or volume or position or assembly of chemical elements, which is located in the middle of the maximum, minimum and / or average size of the nanoparticle, such as half of the diameter of a spherical nanoparticle, or half of the maximum, minimum and / or average length of the nanoparticle. In some other cases, the surface of the nanoparticle is a region or position or assembly of chemical elements, which maintains the maximum distance from the center of the nanoparticle while remaining within the nanoparticle.

[0464] In another embodiment of the present invention, the iron oxide and / or impurities are located at a certain distance from the nanoparticles, preferably at a certain distance from the center or surface of the nanoparticles, greater than 0.001, 0.01, 0.1, 1, 10, 100, 10 3 , 10 5 , 10 10 , 10 20 or 10 50 nm.

[0465] In another embodiment of the present invention, the nanoparticles according to the present invention comprise a core and / or a coating, which preferably surrounds the core of the nanoparticles.

[0466] In one embodiment of the present invention, the core and / or coating of the nanoparticles has at least one property in common with the nanoparticles, such as the concentration of iron oxide and / or impurities.

[0467] In one embodiment of the present invention, the nanoparticles, the core of the nanoparticles and / or the coating have at least one of the following properties:

[0468] (a) Magnetic, diamagnetic, superparamagnetic, ferromagnetic, antiferromagnetic and / or paramagnetic behavior or properties, preferably in an applied magnetic field of intensity greater than 10 -50 、10 -40 、10 -20 、10 -10 、10 -5 、10 -2 or 10 -1 T, preferably observed at a temperature of 10 10 、10 5 、10 3 、10 2 、10 or 1 K. In some cases, the core and the coating may have different magnetic properties. For example, the core may be ferromagnetic or superparamagnetic, while the coating may be diamagnetic or paramagnetic.

[0469] (b) Crystalline part or structure, comprising at least 1, 2, 5, 10, 50, 100, 10 3 、10 5 、10 7 、10 9 、10 20 or 10 50 crystal planes or crystal ordered structures, which can preferably be observed or measured by electron microscopy. In some cases, the core may have a different crystal structure from the coating. For example, the core may comprise more than 1, 5, 10, 10 3 or 10 5 crystal planes or crystal ordered structures, while the coating may have fewer than 10 5 ,10 3 ,10, 5 or 2 crystal planes or crystal ordered structures.

[0470] (c) Composition of a metal or metal oxide, preferably made of iron oxide, most preferably maghemite and / or magnetite. In some cases, the core contains a different composition from the coating. For example, the core contains more than 1%, 5%, 10%, 25%, 50%, 75%, 90%, 95% or 99% by mass of iron oxide, while the coating contains less than 99%, 95%, 90%, 75%, 50%, 10%, 5% or 1% by mass of iron oxide. The percentage can be the ratio between the quantity, volume, number of atoms, mass of iron oxide contained in the core and / or coating and the total quantity, total volume, total number of atoms, total mass of all chemical elements contained in the core and / or coating.

[0471] (d) Single domain, or magnetic monopole,

[0472] (e) A magnetic microstructure, characterized in that there are magnetic field lines that can be oriented to a preferred direction, such as the easy magnetization axis or the crystallization direction of the nanoparticle core, such as

[111] . Such a magnetic microstructure can be observed under certain conditions, especially by electron holography.

[0473] (f) Sizes between 1 nm and 10 5 μm, between 1 nm and 10 3 μm, between 1 nm a and 100 μm, between 1 nm and 10 μm, between 1 nm and 1 μm, between 5 nm and 1 μm, between 5 nm and 500 nm, between 5 nm and 250 nm, between 5 nm and 100 nm, between 5 nm and 80 nm, between 5 nm and 60 nm, between 10 nm and 1 μm, between 10 and 500 nm, between 10 nm and 250 nm, between 10 nm and 100 nm, between 10 nm and 80 nm, between 10 nm and 60 nm, between 15 nm and d 1 μm, between 15 nm and 500 nm, between 15 nm and 250 nm, between 15 nm and 100 nm, between 15 nm and 80 nm, between 15 nm and 60 nm, between 20 nm and 1 μm, between 20 nm and 500 nm, between 20 nm and 250 nm, between 20 nm and 100 nm, between 20 nm and 80 nm, or between 20 nm and 60 nm.

[0474] (g) Sizes greater than 0.1, 1, 2, 5, 10, 15, 20, 25, 30, 35 or 40 nm in certain cases,

[0475] (h) Sizes below 10 10 、10 5 、10 4 、2000、1000、500、400、300、200、150、120、100、95、90、80、75、70、65、60、55、50、45、40、35、30、25、20、15、10 or 5 nm in certain other cases,

[0476] (i) ζ potential, charge or surface charge including between -10 10 mV and 10 10 mV, between -10 5 mV and 10 5 mV, between -10 4 mV and 10 4 mV, between -10 3 mV, -10 2 mV and 10 2mV, between -10 mV and -10 mV. Preferably, the pH value is between 0 and 14, between 1 and 13, between 2 and 12, between 3 and 11, between 4 and 10, between 5 and 9, or between 6 and 8.

[0477] (j) The ζ potential, charge or surface charge, in some cases greater than -10 50 、-10 20 、-10 10 、-10 5 、-10 3 、-10, -5, -1, 0, 5, 10, 20, 50, or 100 mV, preferably at a pH value greater than 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13.

[0478] (k) The ζ potential, charge or surface charge, in some other cases greater than -10 50 、-10 20 、-10 10 、-10 5 、-10 3 、-10, -5, -1, 0, 5, 10, 20, 50 or 100 mV, preferably at a pH value lower than 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 or 0.

[0479] (l) The ζ potential, charge or surface charge, in some other cases lower than 10 50 、10 20 、10 10 、10 5 、10 3 、10, 5, 1, 0, -5, -10, -20, -50 or 100 mV, preferably at a pH value greater than 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13.

[0480] (m) The ζ potential, charge or surface charge, in some other cases lower than 10 50 、10 20 、10 10 、10 5 、10 3 、10, 5, 1, 0, -5, -10, -20, -50 or -100 mV, preferably at a pH value lower than 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 or 0.

[0481] (n) The isoelectric point is between 0 and 14, 1, between 1 and 13, between 2 and 12, between 3 and 11, between 4 and 10, between 5 and 9, or between 6 and 8.

[0482] (o) In some cases, the isoelectric point is greater than 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13 in some other cases, and / or

[0483] (p) In some other cases, the isoelectric point is lower than 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 or 0 in some other cases.

[0484] In one embodiment of the present invention, the core and / or coating are synthesized by cells that generate nanoparticles.

[0485] In another embodiment of the present invention, the core and / or coating are not synthesized by cells that generate nanoparticles.

[0486] In one embodiment of the present invention, the cells that generate nanoparticles (also referred to as cells that generate nanoparticles or (one or more) cells) are eukaryotic cells or prokaryotic cells. In some cases, they are cells generated by, contained in, or amplified in a pre-growth and / or growth medium / culture medium.

[0487] In one embodiment of the present invention, less than 100%, 80%, 70%, 50%, 10%, 20%, 10%, 5%, 2%, 1%, 0.1% or 10 -10 % of the cells that generate nanoparticles contain or generate at least one nanoparticle. In some cases, this percentage can be the ratio between the number of cells contained in a pre-growth and / or growth medium / culture medium that contain or generate at least one nanoparticle and the total number of cells in the pre-growth and / or growth medium / culture medium.

[0488] In another embodiment of the present invention, more than 100, 80, 70, 50, 10, 20, 10, 5, 2, 1, 0.1 or 10-10% of the cells that generate nanoparticles contain or generate at least one nanoparticle.

[0489] In one embodiment of the present invention, the cells that generate nanoparticles are whole cells.

[0490] In another embodiment of the present invention, the cells that generate nanoparticles are part of a cell, such as a cell membrane, vesicle, enzyme, protein, lipid, DNA, RNA, organelle, compartment, cytoplasm, virus, contained in or generated by, originated from, replicated by the generating cell.

[0491] In one embodiment of the present invention, the generating cells are cells that generate nanoparticles, preferably when they are in a growth and / or fed-batch medium, or growing or dividing therein, and avoiding when they are in a pre-production medium, or dividing, growing therein.

[0492] In one embodiment of the present invention, nanoparticles produced by cells are designated as cell-generated nanoparticles.

[0493] In one embodiment of the invention, the cell that produces nanoparticles produces nanoparticles in the cell. The nanoparticles are preferably synthesized in the cell body, when they are generated, assembled, crystallized, partially or completely: i) by a part of the cell or therein or near or inside such as an organelle, Golgi vesicle or its device, endosome, exosome, ribosome, endoplasmic reticulum, actin filament, nucleus, peroxisome, microtubule, lysosome, mitochondria, filament, centrosome, flagellum or cell membrane, ii) located in an area within the cell, or iii) located less than 10 from a part of the cell. 5 ,10 3 ,100,10 or 1nm area.

[0494] In another embodiment of the present invention, the cell producing the nanoparticles produces the nanoparticles extracellularly. The nanoparticles are preferably synthesized outside the cell, when they are produced, assembled, crystallized, partially or completely: i) in an area located outside the cell, or ii) located more than 1, 10, 100, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1000, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000, 10000, 20000, 30000, 40000, 50000, 60000, 70000, 80000, 90000, 10 3 or 10 5 m area.

[0495] In some cases, the cells are higher than 1, 10, 10 3 , 10 5 , 10 10 , 10 20 , 10 50 or 10 100 In some other cases, the cells are less than 10 100 , 10 50 , 10 20 , 10 10 , 10 5 , 10 3 , 100, 50, 10, 5 or 2 cells, preferably per liter of growth medium.

[0496] In one embodiment of the present invention, the nanoparticle producing cell is a eukaryotic cell, preferably belonging to a human, an animal, a plant, a tree, flour, a branch, a mushroom, a fungus, an archaea, a bird, a fish, a pigeon, a trout, a mammal, an ant, a bee or an insect.

[0497] In one embodiment of the present invention, the nanoparticle producing cells are prokaryotic cells or bacteria.

[0498] In some cases, the nanoparticle-producing cells can be mycobacteria, preferably Mycobacterium avium subsp. paratuberculosis, Shewanella, preferably Shewanella oneidensi, Geothrix, preferably Geothrix fermentans. These bacteria preferably synthesize nanoparticles extracellularly.

[0499] In some other cases, the nanoparticle-producing cells can be magnetotactic bacteria, such as Magnetospirillum magneticum strain AMB-1, Magnetococcus marinus strain MC-1, three facultative anaerobic vibrio strains MV-1, MV-2 and MV-4, Magnetospirillum magnetotacticum strain MS-1, Magnetospirillum gryphiswaldense strain MSR-1, Magnetospirillum magneticum strain MGT-1 and the obligate anaerobic bacterium Desulfovibrio magneticus RS-1. These bacteria preferably generate nanoparticles intracellularly.

[0500] In one embodiment of the present invention, the cells that generate nanoparticles are cultured in a pre-growth medium or the pre-growth medium is used during pre-growth, and / or a growth medium is used during growth, and / or a fed-batch medium is used during growth. In some cases, the pre-growth and / or growth medium is a medium in which the nanoparticle-producing cells are amplified. In some cases, the fed-batch medium is a medium added to the growth medium, preferably during the growth step.

[0501] In one embodiment of the present invention, the pre-growth and / or growth medium may contain at least one chemical element, water, and a source of the nanoparticle-producing cells. In some other cases, part of the pre-growth and / or growth medium / medium contains at least one chemical element, a source of water, and does not contain the nanoparticle-producing cells. In some other cases, the pre-growth and / or growth medium only contains the nanoparticle-producing cells.

[0502] In one embodiment of the present invention, the pre-growth and / or growth and / or fed-batch medium contains at least one source, preferably a source of a chemical element, or contains at least one chemical element, preferably in liquid, gaseous and / or solid state. In some cases, the pre-growth and / or growth and / or fed-batch medium is in liquid, gaseous and / or solid state.

[0503] In one embodiment of the present invention, the concentration of a chemical element in the pre-growth and / or growth medium, such as the concentration of iron, is the concentration of the chemical element in: i) the total pre-growth and / or growth medium, ii) a portion of the pre-growth and / or growth medium, or iii) the cells that generate the nanoparticles.

[0504] In one embodiment of the present invention, a quantity or volume of cells can be a quantity or volume of the growth medium containing these cells. In some other cases, a quantity or volume of cells can be a quantity or volume of cells without water or in a non-aqueous cell environment, or the aqueous environment of the cells has been removed, for example, by freeze-drying technology.

[0505] In one embodiment of the present invention, amplifying cells during the pre-growth and / or growth steps can prevent genetic variation of the cells that generate the nanoparticles. In some cases, the genetic variation of the cells that generate the nanoparticles is at least 10 -50 、10 -20 、10 -10 、10 -5 、10 -3 、10 -1 、1, 5, 10, 50, 75, 90 or 95% of the genes, a part of the genes, a DNA part or nucleotide variation. This percentage can be the ratio between the number or quantity of the genes, a part of the genes, a DNA part or nucleotide variation in the cells that generate the nanocell particles and the total number or quantity of the genes, a part of the genes, a DNA part or nucleotide variation in the cells that generate the nanocell particles.

[0506] In another embodiment of the present invention, the pre-growth and / or growth medium contains mostly water, preferably purified water, deionized water or ultrapure water, preferably greater than 10 -50 、10 -20 、10 -10 、10 -5 、10 -3 、1, 10, 50, 75, 80, 90, 99, 99.99 or 99.99999 percentage or mass percentage. This percentage can be the ratio of the quantity, mass, volume or number of atoms of water contained in the pre-growth and / or growth medium divided by the total quantity, mass, volume or number of atoms of all chemical elements contained in the pre-growth and / or growth medium.

[0507] In one embodiment of the present invention, the pre-growth and / or growth medium contains at least one chemical element or a source of a chemical element. In some cases, the concentration of a chemical element (such as iron) in the pre-growth and / or growth medium is the concentration of that chemical element at any time during the pre-growth and / or growth step. In certain cases, this concentration can be measured by estimating the number of moles, mass, or volume occupied by the element divided by the total number of moles, total mass, or total volume occupied by all chemical elements in the pre-growth and / or growth medium.

[0508] In one embodiment of the present invention, the pre-growth and / or growth medium contains at least one carbon source. In some cases, the carbon source includes the chemical elements in the column of the periodic table where carbon is located. In some cases, the carbon source can be selected from the following list: acetate, glycolate, glucose, lactate, pyruvate, succinate, carbon dioxide, glycerol, and derivatives or combinations of these compounds.

[0509] In one embodiment of the present invention, the growth and / or pre-growth medium contains at least one nitrogen source. In certain cases, the nitrogen source includes the chemical elements in the column of the periodic table where nitrogen is located. In certain cases, the nitrogen source can be selected from: ammonium salts, nitrates, urea, amino acids, ammonium salts, ammonia, nitrogen gas, and derivatives or combinations of these compounds.

[0510] In one embodiment of the present invention, the pre-growth and / or growth medium contains at least one sulfur or sulfate source. In some cases, the source of sulfur or sulfate includes the chemical elements in the column of the periodic table where sulfur is located. In some cases, the source of sulfur or sulfate can be sulfate or hydrogen sulfide.

[0511] In one embodiment of the present invention, the pre-growth and / or growth medium contains at least one phosphorus or phosphate source. In some cases, the phosphorus or phosphate source includes the chemical elements in the column of the periodic table where phosphorus is located. In some cases, the phosphorus or phosphate source can be phosphate.

[0512] In one embodiment of the present invention, the pre-growth and / or growth medium contains at least one calcium source. In certain cases, the calcium source includes the chemical elements in the column of the periodic table where calcium is located. In certain cases, the calcium source can be a calcium salt.

[0513] In one embodiment of the present invention, the pre-growth and / or growth medium contains at least one potassium source. In certain cases, the source of potassium includes the chemical elements in the column of the periodic table where potassium is located. In certain cases, the source of potassium is a potassium salt.

[0514] In one embodiment of the present invention, the pre-growth and / or growth medium comprises at least one magnesium source. In some cases, the magnesium source comprises chemical elements in the column of the periodic table where magnesium is located. In some cases, the source of magnesium is a magnesium salt.

[0515] In one embodiment of the present invention, the pre-growth and / or growth medium comprises at least one oxygen source. In some cases, the oxygen source comprises chemical elements in the column of the periodic table where oxygen is located. In some cases, the oxygen source is an organic compound, carbon dioxide or dioxygen.

[0516] In one embodiment of the present invention, the pre-growth and / or growth medium comprises at least one hydrogen source. In some cases, the hydrogen source comprises chemical elements in the column of the periodic table where hydrogen is located. In some cases, the hydrogen source is an organic compound or dihydrogen.

[0517] In one embodiment of the present invention, the pre-growth and / or growth medium comprises at least one iron source. In some cases, the iron source comprises chemical elements in the column of the periodic table where iron is located. In some cases, the iron source is composed of iron or contains iron. In some cases, the source of iron is iron citrate, iron quinolinate, iron chloride or iron sulfate.

[0518] In one embodiment of the present invention, the pre-growth and / or growth medium comprises at least one sulfur source. In some cases, the sulfur source comprises chemical elements in the column of the periodic table where sulfur is located. In some cases, the sulfur source is contained in at least one vitamin.

[0519] In one embodiment of the present invention, the sources of carbon, nitrogen, sulfur, sulfate, phosphorus, phosphate, calcium, potassium, magnesium, oxygen, hydrogen or iron contain greater than 10 -100 、10 -50 、10 -20 、10 -10 、10 -5 、10 -3 、10 -1 、1、5、10、25、50、75、80、90 or 95 mass percentages of carbon, nitrogen, sulfur, sulfate, phosphorus, phosphate, calcium, potassium, magnesium, oxygen, hydrogen or iron. In some cases, they are in a gaseous, liquid or solid state. In some other cases, they can be used to prepare the pre-growth and / or growth medium. In some cases, the pre-growth and / or growth medium contains more than 2, 3, 4, 5, 10, 50, 100, 10 3 、10 5 、10 10 or 10 50carbons, nitrogens, sulfurs, sulfates, phosphors, phosphates, calciums, potassiums, magnésiums, oxygens, hydrogens and / or irons from different sources. In some other cases, the pre-growth and / or growth medium contains less than 2, 3, 4, 5, 10, 50, 100, 10 3 、10 5 、10 10 or 10 50 carbons, nitrogens, sulfurs, sulfates, phosphors, phosphates, calciums, potassiums, magnésiums, oxygens, hydrogens and / or irons from different sources.

[0520] In one embodiment of the present invention, the source of at least one of carbon, nitrogen, sulfur, sulfate, phosphorus, phosphate, calcium, potassium, magnesium, oxygen, hydrogen and / or iron in the pre-growth medium is the same as that in the growth medium.

[0521] In another embodiment of the present invention, the source of at least one of carbon, nitrogen, sulfur, sulfate, phosphorus, phosphate, calcium, potassium, magnesium, oxygen, hydrogen and / or iron in the pre-growth medium is different from that in the growth medium.

[0522] In another embodiment of the present invention, the concentration of the source of carbon, nitrogen, sulfur, sulfate, phosphorus, phosphate, calcium, potassium, magnesium, oxygen, hydrogen and / or iron contained in the pre-growth and / or growth medium is greater than 10 -100 、10 -50 、10 -20 、10 -10 、10 -5 、10 -3 、10 -1 、1、10、10 2 、10 3 、10 5 or 10 10 mM.

[0523] In another embodiment of the present invention, the concentration of the source of carbon, nitrogen, sulfur, sulfate, phosphorus, phosphate, calcium, potassium, magnesium, oxygen, hydrogen and / or iron contained in the pre-growth and / or growth medium is less than 10 100 、10 50 、10 20 、10 10 、10 5 、10 3 、10、1、10 -1 、10 -3 、10 -6 、10 -9 、10 -20 、10 -50 or 10 -100 mM.

[0524] In another embodiment of the present invention, the pre-growth and / or growth medium is prepared using pharmaceutical-grade or ultrapure chemicals or chemical elements.

[0525] In another embodiment of the present invention, the medium impurities are impurities contained in the pre-growth and / or growth and / or fed-batch medium.

[0526] In one embodiment of the present invention, the pre-growth and / or growth medium contains a small amount of medium impurities. In some cases, the percentage content of the medium impurities is less than 100, 10 20 、10 10 、10 5 、10 2 、10, 5, 1, 0.1 or 0.001%. Preferably, the quantity or concentration of the medium impurities contained in the pre-growth and / or growth medium is less than at least 1.00001, 1.1, 1.5, 2, 5, 10, 10 3 ,10 10 or 10 20 times that of the quantity or concentration of at least one source of carbon, nitrogen, sulfur, sulfate, phosphorus, phosphate, calcium, potassium, magnesium, oxygen, hydrogen and / or iron. According to the present invention, in some cases, the percentage of the medium impurities can be defined as the ratio between the number of atoms, quantity, mass or volume of the medium impurities contained in the pre-growth and / or growth medium divided by the total number of atoms, quantity, mass or volume of all chemical elements contained in the pre-growth and / or growth medium. In some other cases, the concentration of the medium impurities contained in the pre-growth and / or growth medium is less than 10 50 、10 30 、10 10 、10 5 、10 3 、500、100、50、10、1、10 -1 、10 -3 、10 -5 、10 -10 or 10 -50 micrograms per milliliter of the pre-growth and / or growth medium..

[0527] In yet another embodiment of the present invention, the pre-growth and / or growth medium / medium contains at least one medium impurity.

[0528] In one embodiment of the present invention, the pre-growth and / or growth medium contains a significant amount of medium impurities. In certain cases, the percentage of the medium impurities (preferably by mass) is greater than 10 -40 、10 -20 、10 -10 、10 -5 、10 -2 、10-1 , 1, 5, 10, 25, 50, 75, 80 or 90%. In some other cases, the concentration of culture medium impurities contained in the pre-growth and / or growth medium is greater than 10 -100 , 10 -50 , 10 -20 , 10 -10 , 10 -5 , 10 -3 , 10 -2 , 10 -1 , 1, 10, 50, 100, 10 3 , 10 5 , or 10 10 micrograms per milliliter of pre-growth and / or growth medium.

[0529] In some cases, the nanoparticles produced or obtained or acquired from the pre-growth and / or growth medium / culture medium can be magnetosomes.

[0530] In one embodiment of the present invention, the method according to the present invention includes a pre-growth step, which includes amplifying cells that generate nanoparticles in a pre-growth medium, so that the cells that generate nanoparticles basically do not generate nanoparticles.

[0531] In one embodiment of the present invention, the cells that generate nanoparticles for starting the pre-growth step are cells having at least one of the following properties: i), they are the cells before the pre-growth step, preferably, 0.001, 0.1, 1, 5, 10, 10 higher than before the start of the pre-growth step 3 , 10 5 or 10 10 hours, ii) they are contained in a cell bank, such as a master cell bank, a working cell bank or a research cell bank, iii) they include more than 1, 5, 10, 10 3 , 10 5 or 10 10 nanoparticles per cell, iv) they are contained in a liquid or culture medium, preferably the same or similar in composition to the pre-growth and / or growth medium, preferably containing a large amount of water, v) they are contained in a medium with a culture medium impurity concentration lower than 100, 10, 1, 10 -1 , 10 -2 , 10 -3 , 10 -5 or 10 -10 μM, vi) they are contained in or maintained under conditions where the medium can maintain or has less than 100, 10, 1, 0.1 or 0.01 grams of impurities per gram of nanoparticles, v) they include between 10 -100 and 10 100 , 10 -50and 10 50 , 10 -30 and 10 30 , 10 -20 and 10 20 , 10 -10 and 101 0 , 10 -6 and 10 5 , 10 -6 and 10 4 , 10 -6 and 10 2 , or 10 -6 and between 1 liter in volume, vi), their volume is at least 10 times smaller than the volume of the first pre-growth step, vii) their cell number, preferably containing between 1 and 10 100 , 2 to 10 50 , 3 to 10 20 , or between 10 and 10 10 cells, vii), their optical density is between 10 -50 to 10 50 , 10 -20 to 10 20 , 10 -10 to 10, 10 -5 to 10 5 , 10 -5 to 10 3 , 10 -5 to 10 2 , 10 -5 to 1, 10 -5 to 10 -1 , 10 -5 to 10 -2 , or 10 -5 to 10 -3 between, viii), they have a certain number of cell divisions, preferably less than 1, 10, 10 3 , 10 5 , 10 10 , 10 20 , 10 50 or 10 100 times per hour or per liter of pre-growth and / or growth medium per hour, ix) they are stored or maintained at a temperature below 100, 50, 25 or 0 °C, preferably 77 K or -20 °C.

[0532] In one embodiment of the present invention, the cells for generating nanoparticles used to initiate the pre-growth step have at least one of the following properties: i) they are contained in a culture medium with an impurity concentration greater than 10 -50 , 10 -20 , 10-10 , 10 -5 , 10 -2 , 10 -1 , 1 or 10 μM of the culture medium, ii), they are contained in a culture medium that can maintain or has more than 10 -40 , 10 -20 or 10 -10 grams of impurities per gram of nanoparticles in the culture medium or are maintained under such conditions, iii), they have a certain number of cell divisions, preferably more than 1, 10, 10 3 , 10 5 , 10 10 , 10 20 , 10 50 or 10 100 , iv) each of them contains less than 1, 5, 10, 10 3 , 10 5 or 10 10 nanoparticles. This situation can occur when / if these cells are maintained in or derived from a culture medium containing a low enough iron concentration to prevent the production of nanoparticles.

[0533] In one embodiment of the present invention, at least one property of the cells for generating nanoparticles used to start the pre-growth step can prevent the death, destruction, disappearance, denaturation, or inactivation of the cells for generating nanoparticles.

[0534] In one embodiment of the present invention, the optical density is measured in the pre-growth and / or growth culture medium, solution, or water. Preferably, the growth culture medium is removed and the cells are redissolved in water. In some cases, the optical density of the cells is measured at a wavelength greater than 1, 2, 5, 10, 50, 100, 200, 300, 400, 450, 500, 550, 600, 900, 10 3 , 10 5 or 10 7 nm. In some other cases, the optical density of the cells is measured at a wavelength lower than 10 7 , 10 5 , 10 3 , 900, 600, 550, 500, 450, 400, 300, 200, 100, 50, 10, 5, 2, or 1 nm. In some other cases, the optical density of the cells is measured at a wavelength between 1 and 10 7 nm, 50 to 10 5 nm, 100 to 10 3 nm, 200 to 900 nm, or 400 to 800 nm.

[0535] In one embodiment of the present invention, the number of cell amplifications between two time points t0 and t1 during the pre-growth and / or growth steps is equal to or proportional to: i) the ratio between the optical density measured at t1 and at t0 and / or ii) the ratio between the number of cells n(t1) at t1 and the number of cells n(t0) at t0.

[0536] In one embodiment of the present invention, the speed or rate of cell division is [n(t1) - n(t0)] / (t1 - t0).

[0537] In another embodiment, the speed or rate of cell division is: [n(t1) - n(t0)] / (t1 - t0), where V is the volume of the pre-growth and / or growth medium in which the cells are cultured or amplified.

[0538] In one embodiment of the present invention, the pre-growth step starts by thawing or heating, preferably from a temperature below 100, 50, 25, 10 or 0 °C to a temperature above 0, 10, 25, 50 or 100 °C, for the cells that generate nanoparticles to start the pre-growth step. After that, the cells that generate nanoparticles are filled or added to the pre-growth medium. In some cases, the initial stage of the pre-growth step occurs between 10 -50 to 10 50 、10 -50 to 10 10 、10 -30 to 10 5 、10 -20 to 10 3 、10 -10 to 10 2 、or 10 -5 to 10 hours.

[0539] In another embodiment of the present invention, the pre-growth step is divided into sub-steps 0, 1,..., i,..., j, corresponding to amplification in different, preferably increasing, volumes, volumes V0, V1, …, V i ,…,,V j 、where i is an integer representing the i-th amplification in different volumes (0 < i < j), j is an integer representing the total number of amplifications in different volumes, V0, Vi and Vj are the initial value, i th and the final amplification volume. In some cases, the cell amplification or the number of cell amplifications in different volumes may be very high during the pre-growth step, for example when the pre-growth step starts from a small number of cells, preferably less than 10 100 、10 50 、10 20 、10 10 、10 5 、103 , 10 2 , 10, 5, 3 or 2 cells, preferably contained in 1 liter or 1 milliliter or 1 microliter of pre-growth medium or aqueous solution. In these cases, i and / or j can be greater than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 10, 10 3 , 10 5 or 10 10 . In some other cases, cell expansion or the number of cells expanded in different volumes may be very low, for example when the pre-growth step starts with a large number of cells, preferably greater than, 3, 5, 10, 10 2 , 10 3 , 10 5 , 10 10 , 10 20 , 10 50 or 10 100 cells, preferably contained in 1 liter or 1 milliliter or 1 microliter of pre-growth medium or aqueous solution. In these cases, i and / or j is less than 10 10 , 10 5 , 10 3 , 10 2 , 10, 5, 4, 3, 2 or 1.

[0540] In one embodiment of the present invention, the ratio Vi / V i-1 is large, preferably greater than 10 -50 , 10 -30 , 10 -20 , 10 -10 , 10 -5 , 10 -3 , 10 -2 , 10 -1 , 1, 1.00001, 1.0001, 1.001, 1.01, 1.1, 1.2, 1.5, 2, 3, 4, 5, 7, 10, 10 2 , 10 3 , 10 5 or 10 10 . In some cases, Vi / V i-1 is large when the number of cell divisions (preferably per hour or per liter of pre-growth medium per hour) between sub-step i-1 and sub-step i of the pre-growth step is greater than 1, 5, 10, 10 3 , 10 10 or 10 20 or when the optical density of the cells increases by more than 1.00001, 1.1, 2, 5, 10, 10 3 , 10 5 or 10 7 times per hour.

[0541] In another embodiment of the present invention, the ratio Vi / V i-1 is low, preferably less than 10 100 、10 50 、10 10 、10 5 、10 3 、10 2 、10, 5, 3, 2, 1.01, 1.001, 1.000001, 1, 10 -5 、10 -10 or 10 -50 。In some cases, Vi / V i-1 is small when the number of cell divisions (preferably per hour or per liter of pre-growth medium per hour) is less than 10 50 、10 10 、10 3 、10 2 、10, 5 or when the optical density of the cells increases by less than 1.00001, 1.1, 2, 5, 10, 10 3 、10 5 or 10 7 times per hour.

[0542] In some cases, the number of pre-growth steps in different volumes can be increased by reducing Vi / Vi-1. In some other cases, the number of pre-growth steps in different volumes can be reduced by increasing Vi / Vi-1.

[0543] In one embodiment of the present invention, the pre-growth step and / or at least one of its sub-steps continue and / or occur until the optical density of the bacterial suspension in volume Vi (1 < i < j) reaches a value of: i) higher than 10 -50 、10 -30 、10 -20 、10 -10 、10 -5 、10 -3 、10 -1 、1, 5, 10, 15, 50, 10 2 、10 3 or 10 5 and / or ii) greater than and preferably greater than 1.01, 1.1, 1.5, 2, 5, 10, 10 2 、10 3 、10 5 、10 10 or 10 20 times, at the end of the pre-growth step compared to the start of the pre-growth step and / or at the end of the pre-growth step compared to at least one sub-step in the pre-growth step.

[0544] In one embodiment of the present invention, the pre-growth step and / or at least one of its sub-steps continue and / or occur until the optical density of the cell suspension in volume Vi (1 < i < j) reaches a value that is: i) less than 10 -50 、10 -30 、10 -20 、10 -10 、10 -5 、10 -3 、10 -1 、1、5、10、15、50、10 2 、10 3 or 10 5 , and / or ii) less than and preferably less than 1.000001, 1.0001, 1.01, 1.1, 1.5, 2, 5, 10, 10 2 、10 3 、10 5 、10 10 or 10 20 times, at the end of the pre-growth step compared to the start of the pre-growth step and / or at the end of the pre-growth step compared to at least one sub-step during the pre-growth step.

[0545] In one embodiment of the present invention, the amplification in volume V0 starts at time point t PG0b , the amplification in volume V0 ends at time point t PG0e , the amplification in volume V i starts at time point t PGib , the amplification in volume Vi ends at time point t PGie , the amplification in volume V j starts at time point t jb , and / or the amplification in volume V j ends at time point t PGje .

[0546] In one embodiment of the present invention, the time length: i) separating the start of the pre-growth step, at time point t PG0b , and the end of the pre-growth step, at time point t PGje , is equal to t PGje - t PG0b and / or ii) separating the start of sub-step i, t PGib , and the end of sub-step i, t PGie , is equal to t PGie - t PGib , greater than or equal to 10 -20 、10 -10 、10 -5 、10 -3 、10 -2 、10-1 1, 2, 5, 10, 24, 100, 10 3 10 5 or 10 7 hours. In some cases, t PGje -t PG0 and / or t PGie -t PGib is larger When the cells are difficult or slow to divide, preferably at less than 10 -50 10 -20 10 -10 10 -5 10 -3 10 -1 1, 10, 10 3 or 10 5 cells divide per hour or at a rate of cells dividing per hour per liter of pre-growth medium.

[0547] In another embodiment of the present invention, t PGje -t PG0b and / or t PGie -t PGib is less than or equal to 10 40 10 30 10 20 10 10 10 5 10 3 10 2 10, 5, 2, 1, 10 -1 10 -3 10 -5 or 10 -10 hours. In some cases, when the cells are easy or fast to divide, t PGje -t PG0b and / or t PGie -t PGib is low, preferably at greater than 10 -50 10 -20 10 -10 10 -5 10 -3 10 -1 1, 10, 10 3 or 10 5 cells divide per hour or at a rate of cells dividing per hour per liter of pre-growth medium.

[0548] In one embodiment of the present invention, the cells generating the nanoparticles are amplified in the pre-growth step by amplifying in t PGib or t PG0bIt is introduced at this point to achieve it or by maintaining the concentration of iron in the pre-growth medium below 10 during the pre-growth step or at least one of its sub-steps 100 、10 20 、10 5 、10 3 、10、5、1、10 -1 、10 -3 or 10 -5 μM to achieve, preferably avoiding the synthesis of nanoparticles that can prevent cell expansion.

[0549] In another embodiment of the present invention, the amplification of the cells generating the nanoparticles in the pre-growth step is achieved by introducing it at t PGib or t PG0b It is introduced at this point to achieve it or by maintaining the concentration of iron in the pre-growth medium greater than 10 during the pre-growth step or at least one of its sub-steps 100 、10 20 、10 5 、10 3 、10、5、1、10 -1 、10 -3 or 10 -5 μM to achieve, preferably enabling effective cell metabolism.

[0550] In another embodiment of the present invention, the amplification of the cells generating the nanoparticles in the pre-growth step is achieved by introducing it at t PGib or t PG0b It is introduced at this point to achieve it or by maintaining the concentration of iron in the pre-growth medium between 10 -10 and 10 10 、10 -5 and 10 5 、10 -3 and 10 3 、10 -1 and 1 μM, 10 -1 and 10 μM, or 10 -2 and 100 μM.

[0551] In another embodiment of the present invention, the nanoparticles-producing cells are amplified in the pre-growth step or at least one of its sub-steps by consuming oxygen. In some cases, the percentage of oxygen in the pre-growth medium starts to decrease from the following values: i) at tPGib or tPG0b, higher than 10 -50 、10 -10 、10 -5 、10 -3Values of 1, 5, 10, 20, 50, 75, 90, 95, 99 or 99.9%, preferably starting from 21% or values between 10% and 30%, at t PGie or t PGje , at least lower than 99.9, 95, 90, 80, 75, 50, 20, 5, 2, 1, 10 -3 , 10 -5 , 10 -10 or 10 -50 %, preferably 0% or values between 10% and 30%, and / or ii) the percentage of oxygen in the pre-growth medium is reduced by more than 1.0001, 1.001, 1.1, 1.2, 1.5, 2, 5, 10, 50, 10 2 , 10 3 , 10 5 or 10 10 times, preferably between t PG0b and t PGje and / or between t PGib and t PGie . In some cases, no oxygen is added to the pre-growth medium during the pre-growth step or at least one of its sub-steps, resulting in a decrease in the percentage of oxygen in the pre-growth medium due to the consumption of oxygen by bacteria. In some other cases, oxygen is added to the pre-growth medium during the pre-growth step or at least one of its sub-steps, resulting in a change in the percentage of oxygen due to the consumption of oxygen by bacteria and the addition of oxygen to the pre-growth medium.

[0552] In one embodiment of the present invention, the percentage of oxygen, preferably the percentage of dissolved oxygen in the pre-growth and / or growth medium, oxygen is preferably in the pre-growth and / or growth medium. In some cases, a percentage of 100% may correspond to the maximum amount of oxygen dissolved in the pre-growth and / or growth medium, including between 10 -5 and 10 20 mg, mainly with 1 to 10 mg of dissolved oxygen per liter of pre-growth and / or growth medium.

[0553] In one embodiment of the present invention, the pre-growth step includes amplifying cells, corresponding to, or being: i) the speed or rate or number of cell divisions, preferably per unit volume, such as 1 liter of pre-growth medium, preferably at the start of the pre-growth step compared to the pre-growth step, one of the sub-steps of the pre-growth step ends, or at t PGie or tPGje is greater than 1.000001, 1.0001, 1.01, 1.1, 1.2, 1.5, 2, 5, 10, 10 PGib or t PG0b than at t 2 , 10 3, 10 5 , 10 10 , 10 20 or 10 50 -fold, ii) the speed or rate or number of cell divisions, preferably per unit volume, e.g., in 1 liter of pre-growth medium, the number of cells or the number of cells per hour is from less than or equal to 10 100 , 10 50 , 10 20 , 10 10 , 10 5 , 10 3 , 10 2 , 10, 5 or 2 at the start of pre-growth or at the start of one of the sub-steps or at t PGib or t PG0b is increased to greater than or equal to 2, 5, 10, 10 2 , 10 3 , 10 5 , 10 10 , 10 20 , 10 50 or 10 100 at the end of pre-growth or at the end of one of the sub-steps or t PGie or tPGje or iii) the optical density, preferably measuring the cells contained in a fixed pre-growth volume (e.g., 1 liter), preferably at the end of the pre-growth step or one of its sub-steps or at tPGie or tPGje is 1.00001, 1.1, 1.2, 1.5, 2, 5, 10, 10 3 , 10 5 , 10 10 , 10 20 or 10 50 -fold, or the optical density increases from less than or equal to 10, 1, 10 -1 , 10 -2 or 10 -3 at the start of the pre-growth step or one of its sub-steps or at tPGib or tPG0b to an optical density greater than or equal to 10 -10 , 10 -2 , 10 -1 , 1 or 10 at the end of the pre-growth step or one of its sub-steps or at t PGie or t PGje .

[0554] In one embodiment of the present invention, preferably during the pre-growth step, at the start, end or at least one of its sub-steps, cells that substantially do not generate nanoparticles have or are characterized by at least one of the following properties i) the nanoparticles contained in the cells are less than 103 ; 10 2 , 50, 20, 10, 5, 2 or 1, preferably less than 10 or 5, or between 0 and 10 3 preferably between 0 and 10 or 0 and 5, ii) a certain percentage of cells having at least one nanoparticle is less than 100%, 99%, 90%, 80%, 50%, 20%, 10%, 1%, 0.1%, preferably less than 10% or 1%, or between 0% and 99%, 0% and 50%, 0% and 10%, preferably between 0% and 5%, where the percentage is preferably the ratio between the number of cells having at least one nanoparticle and the total number of cells preferably contained in the pre-growth medium iii) the optical density is greater than 10 -50 , 10 -20 , 10 -10 , 10 -5 , 10 -3 , 10 -2 , 0.1, 0.2, 0.5, 1, 5, 10 or 100, iv) the number of cells is greater than 1, 5, 10, 10 2 , 10 3 , 10 5 , 10 10 , 10 20 , 10 50 or 10 100 , v) their volume is contained in greater than 0.0001, 0.001, 0.1, 1, 10, 50, 10 2 , 10 3 , 10 5 or 10 10 liters, vi) the number of cell generations is included between 1 and 10 10 , 1 and 10 3 preferably between 50 and 300 of vii) the optical density OD PGE measured at the end of the pre-growth step, and the optical density OD PGB measured at the start of the pre-growth step, the ratio OD PGE / OD PGB , is greater than 1, 2, 5, 10, 15, 25, 50, 100, 10 3 , 10 5 , 10 10 , 10 50 or 10 100 , and / or viii) the optical density OD PGiE measured at the end of sub-step i of the pre-growth step and the optical density OD PGiB measured at the start of sub-step i of the pre-growth step, the ratio OD PGiE / OD PGiB, greater than 1, 2, 5, 10, 15, 25, 50, 100, 10 3 , 10 5 , 10 10 , 10 50 or 10 100 , and / or

[0555] In one embodiment of the present invention, the cells that substantially do not generate nanoparticles are non-magnetic cells.

[0556] In one embodiment of the present invention, preferably at the beginning or end of the pre-growth step or at least one of its sub-steps, a certain percentage of non-magnetic cells is greater than 10 -50 , 10 -20 , 10 -10 , 10 -5 , 10 -3 , 10 -1 , 1, 5, 10, 50 or 75% are obtained. In some cases, the percentage of non-magnetic cells is based on the ratio n NMC / (n MC +n NMC ), where n MC and n NMC are the numbers of magnetic cells and non-magnetic cells, respectively.

[0557] In another embodiment of the present invention, the non-magnetic cells do not exhibit a magnetic response, where the magnetic response can be the orientation of at least one unit parallel to the magnetic field or the cell's migration speed is proportional to the magnetic field strength, where the magnetic field strength can be greater than 10 -9 , 10 -3 , 10 -1 , 1, 10 3 or 10 6 mT and / or the magnetic field is preferably applied to the cells.

[0558] In one embodiment of the present invention, the method includes a growth step that includes amplifying the nanoparticle-generating cells derived from the pre-growth step in a growth medium such that the nanoparticle-generating cells generate nanoparticles. In some cases, the growth step is carried out in a fermenter or device where the temperature, pH, iron concentration, and / or oxygen concentration of the growth medium can be controlled.

[0559] In another embodiment of the present invention, the growth step begins with placing the cells obtained from the pre-growth step into the growth medium. In some cases, the growth step or at least one of its sub-steps is carried out at 10 -50 and 10 50 , 10 -50 and 10 10 , 10 -30 and 105 , 10 -20 and 10 3 , 10 -10 and 10 2 , or occurs within a time period between 10 -5 and 24 hours. In some other cases, the growth step or at least one of its sub-steps occurs below 10 20 , 10 10 , 10 5 , 10 3 , 10 2 , 10, 5, 2, 1, 10 -1 , 10 -2 , 10 -3 , 10 -5 , 10 -10 or 10 -20 . In some other cases, at least one of the growth step or its sub-steps occurs greater than 10 -20 , 10 -10 , 10 -5 , 10 -3 , 10 -2 , 10 -1 , 1, 2, 5, 10, 10 2 , 10 3 , 10 5 , 10 10 or 10 20 .

[0560] In one embodiment of the present invention, the growth step includes amplifying cells during consecutive sub-steps GS O …GS i …GS j wherein t GSOb , t GSib , t GSjb are the start of steps 0, i and j, and t GSOe , t GSie , t GSje are the end of steps 0, i and j, where 0 < i < j. In some cases, i and / or j are greater than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 10, 10 3 , 10 5 or 10 10 . In some other cases, i and / or j are less than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 10, 10 3 , 10 5 or 10 10 .

[0561] In one embodiment of the present invention, each subset i comprises bubbling into the growth medium and / or introducing different amounts of oxygen into the growth medium and / or introducing different amounts of iron into the growth medium, preferably with the aid of a fed-batch medium.

[0562] In one embodiment of the present invention, the growth step comprises at least one of the following sub-steps, during which a gas, such as compressed air or a gas containing more than 1% O2, is introduced into the growth medium, preferably under stirring conditions of 1 to 10 10 , 5 to 10 5 , 10 to 10 4 , 100 to 10 3 or 100 to 300 revolutions per minute, and wherein:

[0563] - During a first sub-step lasting 10 -3 to 10 3 hours or preferably lasting 2 to 16 hours, the flow rate of the gas is between 0 and 10 10 , preferably between 0.001 and 40 millilitres per minute per litre of growth medium, such that the optical density of the cells increases from a value between 10 -10 and 10 3 , preferably between 0.08 and 0.12, at the start of the first sub-step to a value at the end of the first sub-step that is 1.0000001, 1.1, 1.5, 2, 5, 10 or 10 3 times or greater than 10 -9 and 10 4 , preferably between 0.2 and 1.

[0564] - During a second sub-step lasting 10 -3 to 10 3 hours or preferably lasting 2 to 120 hours, the flow rate of the gas is increased by 1.0000001, 1.1, 1.5, 2, 5, 10 or 10 3 times or is between 0 and 10 10 , preferably between 1 and 50 millilitres per minute per litre of growth medium, such that the value of the optical density of the cells increases from the value at the start of the second sub-step, which is equal to the value at the end of the first sub-step or is included between 10 -9 and 10 4 , preferably between 0.2 and 1, to a value at the end of the second sub-step that is 1.0000001, 1.1, 1.5, 2, 5, 10 or 10 3 times or a value between 10 -9 and 10 4 , preferably included between 0.5 and 4.

[0565] - During a third sub-step lasting 10 -3 to 10 3 hours or preferably 2 to 120 hours, the flow rate of the gas is increased by 1.0000001, 1.1, 1.5, 2, 5, 10 or 10 3 times or between 0 and 10 10 inclusive, preferably between 50 and 120 milliliters per minute per liter of growth medium, such that the optical density of the cells increases from the start of the third sub-step, which is equal to the end of the second sub-step, or is included between 10 -9 and 10 4 inclusive, preferably between 0.5 and 4, to a value at the end of the third sub-step that is 1.0000001, 1.1, 1.5, 2, 5, 10 or 10 3 times or a value between 10 -9 and 10 4 inclusive, preferably included between 1 and 8 at the end of the third sub-step.

[0566] - During a fourth sub-step lasting 10 -3 to 10 3 hours or preferably 2 to 120 hours, the flow rate of the gas is increased by 1.0000001, 1.1, 1.5, 2, 5, 10 or 10 3 times or between 0 and 10 10 inclusive, preferably between 200 and 300 milliliters per minute per liter of growth medium, such that the optical density of the cells increases from the start of the fourth sub-step, which is equal to the end of the third sub-step, or is included between 10 -9 and 10 4 inclusive, preferably between 1 and 8, to a value at the end of the fourth sub-step that is 1.0000001, 1.1, 1.5, 2, 5, 10 or 10 3 times or a value between 10 -9 and 10 4 inclusive, preferably included between 2 and 16 at the end of the fourth sub-step.

[0567] - During a fifth sub-step lasting 10 -3 to 10 3 hours or preferably 2 to 120 hours, the flow rate of the gas is increased by 1.0000001, 1.1, 1.5, 2, 5, 10 or 10 3 times or between 0 and 10 10 inclusive, preferably between 300 and 500 milliliters per minute per liter of growth medium, such that the optical density of the cells increases from the start of the fifth sub-step, which is equal to the end of the fourth sub-step, or is included between 10-9 and 10 4 Between, preferably between 1 and 8, the value at the end of the fourth sub-step is 1.0000001, 1.1, 1.5, 2, 5, 10 or 10 times greater than the value at the start of the fourth sub-step 3 times or 10 -9 and 10 4 The value between, preferably included between 4 and 32 at the end of the fifth sub-step.

[0568] In one embodiment of the invention, during sub-step i, preferably sub-steps 2 to 5: i) the percentage of oxygen is maintained above 0.01% or 0.1 mBar by the air flow and below 0.9% or 9 mBar due to cell oxygen consumption, ii) the gas flow is between 0 and 10 10 、1 and 10 5 、5 and 10 4 、10 and 10 3 ml per minute per liter of growth medium, iii) the growth medium is stirred at a rate of 1 to 10 5 ,10 and 10 4 、50 and 10 3 ,or 100 to 500 revolutions per minute, iv) the gas flow can be reduced by increasing the stirring rate of the medium v) the gas flow can be reduced by decreasing the stirring rate of the medium, and / or vi) the optical density of the cells increases from a value preferably included between 10 -50 and 10 3 at the start of sub-step i to a value preferably included between 10 -20 and 10 5 at the end of sub-step i.

[0569] In one embodiment of the invention, the growth step comprises at least one sub-step in which: i), the pH of the growth medium is maintained at a fixed or determined or selected pH, including between 0 and 14, 2 and 13, 4 and 11, 5 and 10, preferably included between 5 and 8, most preferably equal to 6.9, preferably by adding an acidic iron source included in the fed-batch medium, preferably under stirring conditions of 1 to 10 10 preferably 100 to 300 revolutions per minute. In some cases, the iron concentration contained in the growth medium at the start of the growth step or at one of its sub-steps, i.e.: i), is below 10 10 、10 5 or 10 2 μM, preferably below 10 or 2 μM, and / or ii), is included between 10 -10 and 10 10 、10 -5 and 10 5 、10-3 and 10 3 between 0.2 and 20 μM, preferably between 0.2 and 20 μM. In some other cases, during the growth step or one of its sub-steps, the iron concentration of the growth medium is increased, preferably by adding a fed-batch medium to the growth medium, to reach a value at the end of the growth medium or one of its sub-steps that is:

[0570] i) greater than 10 -10 , 10 -5 , 10 -1 or 1 μM, preferably greater than 2 μM and / or ii) comprised between 10 -10 and 10 10 μM, preferably between 2 μM and 5 mM or between 2 μM and 0.5 mM.

[0571] - In one embodiment of the invention, the growth step comprises at least one of the following sub-steps, wherein: during the first sub-step (lasting from 10 -3 to 10 3 , preferably 2 to 16 hours), a fed-batch medium is added to the growth medium to bring the iron concentration in the growth medium between 10 -10 μM and 10 10 μM. Preferably between 2 and 20 μM, in some cases without considering the iron consumption by the bacteria, and in some other cases taking into account the iron consumption by the bacteria. This preferably results in an increase in the production of nanoparticles from the start to the end of the first sub-step that is higher than 1.0000001, 1.1, 1.5, 2, 5, 10 or 10 3 times or a value between 10 -10 and 10 10 preferably, from 0.001 to 0.1 mg of nanoparticles per liter of growth medium at the start of the first sub-step to a value between 10 -10 to 10 10 preferably, 1 to 10 mg of nanoparticles per liter of growth medium at the end of the first sub-step.

[0572] - During the second sub-step (lasting from 10 -3 to 10 3 , preferably 2 to 120 hours), a fed-batch medium is injected into the growth medium to bring the iron concentration in the growth medium between 10 - 10 to 10 10 μM. A fed-batch medium is added to the growth medium to bring the iron concentration in the growth medium between 10 -10 μM and 10 10Between μM. Preferably between 20 and 40 μM, in some cases without considering the consumption of iron by bacteria, and in some other cases taking into account the consumption of iron by bacteria. This preferably results in an increase in the generation of nanoparticles from the start of the second sub-step to the end of the second sub-step that is higher than 1.0000001, 1.1, 1.5, 2, 5, 10 or 10 3 times or from 10 -10 to 10 10 Values between, preferably, 1 to 10 mg of nanoparticles per liter of growth medium at the start of the second sub-step to 10 -10 to 10 10 Values between, preferably 2 to 20 mg of nanoparticles per liter of growth medium at the end of the second sub-step.

[0573] - During the third sub-step (lasting 10 -3 to 10 3 , preferably 2 to 120 hours), a fed-batch medium is injected into the growth medium so that the iron concentration in the growth medium is between 10 - 10 to 10 10 μM. The fed-batch medium is added to the growth medium so that the iron concentration in the growth medium is between 10 -10 μM and 10 10 μM. Preferably between 40 and 150 μM, in some cases without considering the consumption of iron by bacteria, and in some other cases taking into account the consumption of iron by bacteria. This preferably results in an increase in the generation of nanoparticles from the start of the third sub-step to the end of the third sub-step that is higher than 1.0000001, 1.1, 1.5, 2, 5, 10 or 10 3 times or from 10 -10 to 10 10 Values between, preferably, 2 to 20 mg of nanoparticles per liter of growth medium at the start of the third sub-step to 10 -10 to 10 10 Values between, preferably 4 to 40 mg of nanoparticles per liter of growth medium at the end of the third sub-step.

[0574] - During the fourth sub-step (lasting 10 -3 to 10 3 , preferably 2 to 120 hours), a fed-batch medium is injected into the growth medium so that the iron concentration in the growth medium is between 10 - 10 to 10 10 μM. The fed-batch medium is added to the growth medium so that the iron concentration in the growth medium is between 10 -10 μM and 10 10Between μM. Preferably between 150 and 500 μM, in some cases without considering the bacterial consumption of iron, and in some other cases taking into account the bacterial consumption of iron. This preferably results in an increase in the generation of nanoparticles from the start of the fourth sub-step to the end of the fourth sub-step by more than 1.0000001, 1.1, 1.5, 2, 5, 10 or 10 3 times or from 10 -10 to 10 10 values between, preferably, 4 to 40 mg of nanoparticles per liter of growth medium at the start of the fourth sub-step to 10 -10 to 10 10 values between, preferably 8 to 80 mg of nanoparticles per liter of growth medium at the end of the third sub-step.

[0575] - During the fifth sub-step (lasting from 10 -3 to 10 3 , preferably 2 to 120 hours), a fed-batch medium is injected into the growth medium such that the iron concentration in the growth medium is between 10 - 10 to 10 10 μM. The fed-batch medium is added to the growth medium such that the iron concentration in the growth medium is between 10 -10 μM and 10 10 μM. Preferably between 500 and 1000 μM, in some cases without considering the bacterial consumption of iron, and in some other cases taking into account the bacterial consumption of iron. This preferably results in an increase in the generation of nanoparticles from the start of the fifth sub-step to the end of the fifth sub-step by more than 1.0000001, 1.1, 1.5, 2, 5, 10 or 10 3 times or from 10 -10 to 10 10 values between, preferably, 8 to 80 mg of nanoparticles per liter of growth medium at the start of the fourth sub-step to 10 -10 to 10 10 values between, preferably 16 to 160 mg of nanoparticles per liter of growth medium at the end of the third sub-step.

[0576] In one embodiment of the present invention, during the growth step or sub-step i of the growth step, preferably sub-steps 1 to 5: i) the iron concentration of the growth medium is increased to more than 10 -10 μM, preferably 2 μM, preferably by adding a fed-batch medium containing iron to the growth medium and less than 10 10 mM, preferably 5 mM, due to the cell consumption of iron, ii), the total amount of iron added to the growth medium per liter of growth medium is included between 10 -6 and 15, preferably at 2.10 -4and 1.5 g iron per liter of growth medium, and / or iii) the amount of nanoparticles is increased from a value between 0 and 500 mg, preferably between 0 and 80 mg at the beginning of the growth step or one of its sub-steps to between 1 and 10 mg at the end of the growth step or one of its sub-steps 5 A value between mg, preferably between 10 and 200 mg of nanoparticles per liter of growth medium.

[0577] In one embodiment of the invention, the growth step comprises cell expansion, which is related to or corresponds to the speed or rate or number or optical density of cell division, and preferably is greater than 1.000001, 1.0001, 1.01, 1.1, 1.2, 1.5, 2, 5, 10 ... 2 , 10 3 , 10 5 , 10 10 , 10 20或 10 50 times.

[0578] In one embodiment of the present invention, preferably during the growing step, during its beginning or at its end or at least one sub-step thereof, the cells producing the nanoparticles have at least one of the following properties: i) a number of nanoparticles contained in the cells is greater than 1, 2, 5, 10, 50, 10 2 or 10 3 , preferably greater than 0, 1 or 2, or from 0 to 10 3 between 0 and 100 or between 0 and 10, ii) a certain percentage of cells have at least one nanoparticle greater than 10 -4 ,10 -2, 10 -1 , 1, 5, 10, 50, 75 or 95, preferably greater than 10 or 50% or comprised between 0 and 99%, 10% and 75%, 5% and 90%, preferably between 20% and 100%, iii) an optical density greater than 10 -50 ,10 -20 ,10 -10 ,10 -5 ,10 -3 ,10 -2 , 0.1, 0.2, 0.5, 1, 5, 10 or 100, iv) a certain number of cells greater than 1, 5, 10, 10 2 , 10 3 , 10 5 , 10 10 , 10 20 , 10 50 or 10 100. v) The cells are contained in a volume greater than 0.0001, 0.001, 0.1, 1, 10, 50, 10 2 , 10 3 , 10 5 or 10 10 liters. vi) The number of cells produced is between 1 and 10 10 , 1 and 10 3 , preferably between 50 and 300. vii) The optical density, OD GE measured at the end of the growth step and the optical density OD GB measured at the start of the growth step, the ratio between OD GE / OD GB is greater than 1, 2, 5, 10, 15, 25, 50, 100, 10 3 , 10 5 , 10 10 , 10 50 or 10 100 , or viii) The ratio ODGiE / ODGiB of the optical density OD GiE measured at the end of sub-step i of the growth step to the optical density ODGiB measured at the start of sub-step I, which is greater than 1, 2, 5, 10, 15, 25, 50, 100, 10 3 , 10 5 , 10 10 , 10 50 or 10 100 .

[0579] In one embodiment of the invention, the cells that substantially do not produce nanoparticles are non-magnetic cells.

[0580] In one embodiment of the invention, during the growth step, at the start or end or at least one of its sub-steps, the percentage of magnetic cells obtained is greater than 10 -50 , 10 -20 , 10 -10 , 10 -5 , 10 -3 , 10 -1 , 1, 5, 10, 50 or 75%. In some cases, the percentage of magnetic cells is equal to n MC / (n MC + n NMC ), where n MC and n NMC are the numbers of magnetic cells and non-magnetic cells respectively. In another embodiment of the invention, the magnetic cells are cells that exhibit a magnetic response.

[0581] In some cases, the pre-growth step, the growth step or at least one of its sub-steps is carried out at a temperature greater than -250, -200, -150, -100, -50, -20, -10, -5, -2, -1, 0, 1, 2, 5, 10, 20, 50, 75, 100, 10 3 、10 5 or 10 7 ℃, or the temperature change is greater than 10 -5 、10 -3 、10 -2 、0.1, 1, 5, 10, 50, 100 or 150℃. In some other cases, the pre-growth step, the growth step or at least one of its sub-steps is carried out at a temperature lower than 0 7 、10 5 、10 3 、100, 75, 50, 40, 30, 20, 10, 5, 2, 1, 0, -1, -2, -5, -10, -20, -50, -100, -150, -200 or -250℃, or the temperature change is lower than 10 5 、10 3 、10 2 、50, 20, 10, 5, 2, 1 or 0.1℃.

[0582] In some cases, the pre-growth step, the growth step or at least one of its sub-steps is carried out at a pH greater than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14, or the pH change is greater than 10 -10 、10 -7 、10 -5 、10 -4 、10 -3 、10 -1 、1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 pH units. In some other cases, the pre-growth step, the growth step or at least one of its sub-steps is carried out at a pH lower than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 or the pH change is lower than 10 -10 、10 -7 、10 -5 、10 -4 、10 -3 、10 -1 、1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 pH units.

[0583] In some cases, the temperature, the temperature change, the pH or the pH change is large enough such that at least 1, 5, 10, 10 3 、105 , 10 10 or 10 50 cell divisions or cell divisions per hour.

[0584] In some other cases, the temperature, temperature change, pH or pH change is low enough to prevent more than 1, 5, 10, 10 3 , 10 5 , 10 10 or 10 50 cell destruction, disappearance or denaturation per hour.

[0585] The invention also relates to a method according to the invention, wherein the pre-growth medium does not contain iron or at least one iron source.

[0586] The invention relates to a method according to the invention, wherein the pre-growth medium contains iron or at least one iron source, and the nature and / or quantity of the iron or iron source preferably do not allow nanoparticles to be generated and cell growth to proceed simultaneously.

[0587] In one embodiment of the invention, the nature of the iron source is the composition, chemical formula, type of iron source, or the iron source itself. In some cases, the iron source is a source of iron(III) or iron(II). In some cases, it can be or contain or be made of or have the chemical formula Cl3Fe, C 10 H 12 FeN2NaO8, Fe2O 12 S3, C6H8FeNO7, C6H5FeO7, FeH 18 N3O 18 , C 30 H 21 FeN3O 15 -3 , FeO4P, C6H7FeO8, Fe2H2O 13 S3, Fe2H 12 O 18 S3, C 10 H 12 FeN2NaO8, C 10 H 13 FeN2O8, FeH 28 NO 20 S2, C 10 H 15 FeN2NaO8, C 10 H 14 FeN2NaO8 +4 , C 14 H 21 FeN3O 10 , C 18 Fe7N 18, Fe4H2O 22 S5, Fe4O 21 P6, F3Fe, C6H 11 FeNO7 +3 , C6H 11 FeNO7, C 18 H 15 FeO9, C 12 H 29 Fe5Na2O 23 , C 12 H 22 Fe2O 14 , C 15 H 21 FeO6, C 15 H 24 FeO6, C6H5FeO7, C 10 H 16 FeN3O8, C4H 10 FeO5, C 54 H 105 FeO6, AsFeH 13 O9 + , AsFeO4, Fe +3 , C6H 12 FeN3O 12 , C6H 18 As3FeO6, FeH2O5P, C 21 H 21 FeO9S3, C6H 11 FeNaO7 +3 , C 14 H 22 FeN3NaO 10 , FeNaO7P2, C3H9As3Fe3O9, C 18 H 24 Fe4O 42 P6, C6H 11 FeO 10 , C9H 18 FeN3S6, Cl3FeO 12 , C6H9FeNO7 + , Cr3Fe2O 12 , C6H 10 FeNO8, FeH3O3, C 15 H 30 FeN3S6, C 30 H 27 FeN3O 15 , C3FeN3S3, C6H 12 FeKO6 +4, FeH3O3, FeN3O9, C3H3FeO6, C6H8FeO7, C 24 H 45 FeO6, FeO6P3, Fe2H 14 O 19 S3, C 18 H 33 FeO 21 , C6H9FeO9, C 18 H 27 FeO 24 , C6FeN6 -3 , C 10 H 12 FeN2O8 - , C 22 H 36 N4O 13 , C3FeN3, C6H 12 FeN3O 12 , C6H9FeO6, C 15 H 27 FeO6, FeH4O6P, C 21 H 15 FeO9, FeH8O8P, C6H6FeNO6, C4FeKO8, C 12 H 12 Fe2O 18 , C 33 H 35 FeN4O4, Cl3FeH4O2, C 24 H 45 FeO6, C 10 H 15 FeN2O7, FeH4NO8S2, C 32 H 31 FeN4O5, Fe2H6O3, AlF6Fe, C4H4FeNO8, C 81 H 84 FeN3O 33 , Fe2S3, Cl3FeH 14 O7, C 18 H6FeN9O 21 , Cl3FeO9, FeI3, C6H 14 FeO 10 , C6H 10 FeO8, C 55 H 80 FeN 17 O 21 S3, C 10 H 16 FeN5O 13 P3, C 18 H34 FeO 16 +3 、 C 12 H 12 Fe2O 15 、 C6FeNa3O 12 、 C 10 H 12 FeKN2O8、 C 21 H 24 FeN3O9、 C6H6Fe2O 12 、 C6Fe2O 12 、 AsFe、 C 35 H 33 FeN5O 11 -3 、 Cl3FeH2O、 C 18 H 30 Fe2N6O 12 、 FeI3O9、 C 10 H 18 FeN2NaO 11 、 Cl3FeH 18 O9、 Cr2FeH4NO8、 C9H 21 Fe2O 18 P3、 C 18 H 34 FeO2、 C 30 H 27 FeO6、 C 30 H 24 FeN3O 15 、 C 54 H 102 FeO6、 Fe4H 18 O 30 P6、 Fe2Se3、 C 54 H 99 FeO6、 C 15 H 21 FeO6、 C 10 H 18 FeN2O7 +2 、 C 10 H 18 FeN2O7 +2 、 C 10 H 19 FeN3O8、 C 22 H 14 FeO4、 C 39 H 63 FeN6O 15 +3 、 C 10 H 19 FeN3O8、 C4FeNaO8、 FeO4V、 C6H15 FeN3O 12 、C6Fe2O 12 、C 18 H 24 Fe2O 24 -6 、C 18 H 19 FeN2NaO6、C 18 H 19 FeN2NaO6、C 12 H 18 Fe2O 12 、C6FeK3N6、C 24 H 47 FeO 25 - 、C 18 H 38 FeO 19 、C 15 H 21 FeO6、C 18 H 39 FeO 24 、C6H 11 FeNO7、C6H 12 FeO6、C 12 H 28 FeO 14 、FeHO2、C 45 H 36 FeN3O6、Fe3H2O4、Fe2O3、C 36 H 72 FeO6、C 12 H 18 Fe2O 15 、C9H 18 FeO9、FeH6O3、C 54 H 102 FeO6、C 42 H 84 FeO6、C 16 H 31 FeO2 +2 、C 36 H 69 FeO6、Fe3H8O4、C8H 15 Fe2O2 +4 、C 12 H 48 Fe2N 12 O 12 S3、C 48 H 96 FeO6、C9H 15 FeO9、C 35 H 39N5O 11 、C 42 H 81 FeO6、C 48 H 93 FeO6、C 10 H 24 O2、Fe2H 18 O 21 S3、FeH 12 N3O 15 、C 24 H 23 FeN 10 O6S2、C 18 H 14 Cl3FeN 10 S2、C 21 H 15 FeO6、Fe2H 10 O 17 S3、C 10 H 19 FeN3O8、C 18 H 20 FeN2NaO6、C3F9FeO9S3、C5H 14 FeO4、C6H 19 FeNO 11 、C 18 H 16 FeN2NaO6、C 32 H 36 N4O9、C 15 H 30 FeO6、C 15 H 24 FeO6、C 15 H 15 F9FeO6、C 21 H 21 Cl3FeIS、C6H 12 Fe2O 18 、C6H 18 FeO 12 、C6H 15 FeO 12 、C6H 18 Fe2O 18 、C6H8FeO7、C6H 13 FeO 11 、C6H4Fe2N7、FeH2O4S、C 42 H 60 N 12 O 16 、C6Fe2N6、C3Fe2O9、C 162 H 297 FeO 27-6 , C 21 H 27 Cl4FeN2O, C6H4FeNaO7, C 27 H 50 FeN6O 10 , C 25 H 48 FeN6O8, C 27 H 48 FeN6O9, C6H7FeO6 +2 , Fe2H2O4, C 14 H 26 FeN5O 10 , Cl4FeH4N, Cl3FeH 12 O 18 , C6H 17 FeN2NaO7 +3 , C 10 H 11 , CFeNO6, C 15 H 15 F9FeO6, C6Fe2N6Na, C9H 21 Fe2O 18 , C3P3, C 21 H 27 , CClN2O, C2H3FeO, C 10 H 12 Fe2N2O8, FeH3O3P, C7H5FeO2, C7H5FeO2, FeI3O 12 , C3H4FeNO2S, C2H2FeNO2, C 12 H 12 Fe2O 12 , C8H7FeNO3, C2HFe, C6H7FeO2S4, C6H 11 FeO6, C 14 H 19 FeO 12 , BFeH3O3, C 21 H 18 FeO 15 , C 35 H 56 FeN6O 13 , C 12 H 30 , CFeO3, CHFe, C 47 H 48 FeNO 14 , Fe2H6O3, Fe2O9Sn3, C 18 H 18FeO3, Fe2O9Se3, Fe2O9Si3, Fe2O9S3, Br3FeO9, FeN3O6, C 24 H 54 FeO3, C 66 H 129 FeO6, FeP, C6H 18 FeO 24 P6 +3 , C 33 H 72 FeO3, C 40 H 75 FeO4, C2H3FeS, C3FeN3, C 21 H 39 FeO6, FeSi, C 30 H 29 FeN3O 16 , C 22 H 36 FeN4O 13 , C 30 H 57 FeO6, C 60 H 117 FeO6, C 18 H 12 FeN3O6, C 18 H 31 FeO2 +2 , FeS2, C6H 11 FeN4O2, C6H5FeO7, C6H5FeS, C 10 H 13 FeN2O 10 - , C8H 13 FeOS2, C 27 H 51 FeO6, C 24 H 44 FeO 25 - , C6H 15 FeN3O6, C6H 12 FeO9, Cl3FeO9S3, CFeNS, Fe4H 12 O 12 Si3, C3H6FeO 12 , C4H3FeO4S2, C4H4FeO6, C6H3FeN3O6, C5H5FeO2, C 10 H 24 FeN4O9, C 14 H 19 FeN3NaO 10 , C 10 H14 FeN2Na2O8, C 36 H 44 FeN4, C6FeNa3O 12 , Fe2H3OS3, C 16 H 27 FeO4, C6H8Fe2O 13 , C6H7FeO3, C4H4FeO6S2, C2H5FeN2, C5H7FeOS2, C 18 H 18 FeNa6O 21 , C3H9FeO9S3, C 24 H 54 FeO 12 P3, C 36 H 55 FeN6O 11 , Fe2H2O 10 Si3, C2H4FeNO2, C4H 11 FeN2O4, AsFeH2O5, C 12 H 13 FeO 13 , C 36 H 67 FeO6, C 12 H 13 FeO 13 , C3H6FeN3O6, C 18 H 15 FeO9S3, C 36 H 75 FeO 12 S3, Fe2H4O5, C 28 H 24 FeN4 +3 , F3Fe, C 30 H 30 FeO6, BFe, C2H8N2O4, C8H5FeN2O5, Fe2H4O 11 Se3, C6H7FeO6S4, C4H 10 FeN3, C6H 12 Fe2O 15 , C 15 H 23 FeO5, C8H 12 FeNO 12 , C 49 H 56 ClFeN4O6, FeH4NO8S2, C 36 H 75 FeO9S3, B3F 12 Fe, FeP, Fe2H20 O 22 S3, Cl3FeH 12 O 15 , C 18 H9FeN6, Fe2H 12 O 15 Se3, C 56 H 51 FeN4, Fe2H8O 13 Se3, C 44 H 27 FeN4, C 33 H 30 FeN4O6 -2 , CrFeO3, C 18 H 12 FeN3O 15 S3, Cl3FeH 18 O 21 , C6H5FeNa3O 13 , C 18 H 14 FeN 13 O9S2, C 15 H 24 FeO6, C 24 H 27 FeO9S3, C 27 H 54 FeN3S6, Cl3FeH 12 O6, C 16 H 36 Pb, C8H 18 Fe2O 12 P2, Cl3FeH 24 O 12 , C 24 H 30 FeO9S3, C 21 H 24 FeO9S3, C 18 H 15 FeO 12 S3, Cl3FeH 20 O 10 , C 28 H 24 FeN6O6 + , C 66 H 121 Fe2NaO 65 , Cr3FeH3O 12 , C 12 H 28 Fe2O 14 , C3H8FeNS2Zn - , F3FeH6O3, C30 H 51 FeO6, C 30 H 48 Fe4N6O 24 , C 30 H 18 FeN3O6, C 20 H 36 FeO4, C6H6FeK3O 15 , C 15 H6F 18 FeO6, C 10 H 13 FeN2O8, C6FeN6, C 15 H3F 18 FeO6, C 15 H 12 FeN3O3S3, C 21 H 23 FeO 10 S3, FeH2O +3 , C 24 H 44 FeNaO 28 , Cr3FeO6, Fe2H2O +6 , C6H 12 FeN9, FeH5NO4S, C2K2O4, C 18 H 13 FeN6, C 30 H 27 FeO6, C 34 H 38 N4O4, Cl3FeH 15 O 18 , C6H 18 FeO6P3S6, C6H 11 ClFeNO 10 S2, C5H4F3FeO2 +2 , C6H6Cr2O 12 , C4H3CrKO8, C2MgO4, C 12 H 25 FeO 14 , C2H2MgO4 +2 , C2CrO4 + , C2HNaO4, C2HKO4, C6Cr2O 12 , C2H2FeO4, C2H4MgO6, C6AlO 12 -3 , C6Al2O 12 , C2Li2O4, C2MgO4, C 44 H 30 N4O12 S4, C 10 H 19 FeN2NaO 10 , C5H4CuFeN6O3, C 10 H 14 FeN2NaO9, C 30 H 15 FeN3Na3O 15 S3, C 27 H 15 FeN 12 O6, C9H 18 FeN3S6, C 30 H 30 FeN3O 15 +3 , C9H 18 FeN3S6, C6FeN6, C 18 Fe7N 18 , C 18 H 18 FeN2NaO6, C 30 H 21 FeN 12 O6, C 44 H 30 FeN4 +3 , C 14 H 18 FeK2N3O 10 , C 10 H 16 FeN2NaO8, C 33 H 29 FeNO 11 + , C 25 H 18 FeN4O6S + , C 35 H 24 FeN6O2S + , C 32 H 32 ClFeN4O6, C 30 H 12 F9FeN 12 O6, C 30 H 18 Cl3FeN 12 O9, C 60 H 72 FeN9O9 +3 , C 60 H 66 FeN9O9 +3 , C 15 H 24 FeO6, C22 H 25 Cl2FeN3O9 + ,C 18 H 23 Cl3FeN3O 12 , C 11 H 24 FeNO 11 , C 49 H 54 FeN4O9 + ,C 42 H 54 Cl8Fe2N4O2、C 44 H 26 Cl4FeN4 +3 , C 34 H 32 FeN4O4 + ,C 44 H 38 FeN8 +7 、C9H 11 Cl2FeN4O2S、C 18 H 32 FeN4O8 +3 , C 34 H 32 ClFeN4O6, or C 19 H 25 FeN4O6. In some other cases, the source of iron can be or contain or consist of or have the chemical formula: Fe +2 ,FeH 14 O 11 S, FeH8N2O8S2, FeO4S, Cl2Fe, FeS, C4H2FeO4, C 12 H 26 FeO 16 、C4H5FeNO4、C 12 H 10 Fe3O 14 , C 16 H 30 FeO4, FeH2O5S, C 10 H 12 FeN2Na2O8, As2Fe3O8, CFeO3, C6H 12 FeO6、FeH 12 N2O 12 , C 12 H 10 Fe3O 14 、C6H5FeNaO7、C 34 H 32 FeN4O4、C 12 H22 FeO 14 、 C 12 H 14 FeO 12 、 C6H 10 FeO6, C4H8FeN2O4, C 12 H 28 FeO 16 、 FeI2, FeH4N2O6S2, C 34 H 32 FeN4O4 -2 、 C 34 H 32 FeN4O4, F2Fe, C6H 18 FeO9, C6H5FeO7 - 、 C2FeO4, C4H4FeO4, Cl2FeO8, Fe3O8P2, FeO, B2F8Fe, FeH8O8S, C4H6FeO4, C4H4FeO4, C 12 H 10 FeNa4O 14 、 C 22 H 14 FeO4, C2H4FeO6, C 12 H 24 FeO 14 、 C 14 H 20 FeN3O 10 - 、 Cl2FeH8O4, C 12 H8FeN2O4, C4H8FeO4, C5H7FeNO4, C8H 12 FeN2O8, C 12 H 10 Fe3O 14 、 C6H 16 FeO9, C 19 H 19 FeN7O 10 S, C 10 H 16 FeN2O8, C 12 H 10 Ca2FeO 14 、 C2H6FeO6, C 36 H 70 FeO4, C6H6FeO7, C4H2FeO4, C 36 H 21 Cl2FeN9O 14 、 C 32 H 62FeO4, FeH2O2, C4H6FeO6, C6H8CaFeO7 +4 , C4H 10 Cl2FeN2O4, C 36 H 24 Cl2FeN6O8, C6H 14 FeO7, C 12 H 16 FeO 12 , BFe, C 32 H 16 FeN8, C 12 H 26 FeO 15 , C 12 H 10 Fe3O 14 , FeH8I2O4, C4H 10 FeN2O8S, C 30 H 24 Cl2FeN6O8, C 39 H 30 Cl2FeN6O8, C 12 H 14 FeO 12 , C 30 H 24 FeN6 +2 , C4H2FeO4 -2 , C4H4FeO4, C 10 H 16 FeO4, C 36 H 24 FeN6O4S, C2H4FeO6, C2H2FeO6, C8H 15 Fe2O2 +4 , C 32 H 16 FeN8, C 12 H 16 Fe3O 14 , C 12 H 24 FeO 14 , C2FeN2S2, C 12 H 16 FeN6O4, C 14 H 20 FeN3O 10 , C 12 H7FeN3O6S, C 20 H 12 FeN4, C 12 H 16 Ca2FeO 14 , C 46 H54 FeO9, C6H5FeO7, FeH4O6S, C 10 H 15 FeN2NaO7, C 10 H6FeN4O8, Fe2P, C4H4FeO6, C 14 H 26 FeO 16 , Cl2FeH 12 O 14 , C4H8Cl2FeN2O4, C6Fe3N6, C4H 12 As2FeO8, C 10 H 16 FeO4, FeH 20 N2O 14 S2, C 16 H 30 FeO4, C 40 H 40 FeN8O4 + , Fe2Na8O 21 P6, C 14 H8FeO 10 , C 14 H8FeO4, C 12 H 20 FeO4, C8H8FeS, C5H4FeO, C2H3FeNO2, C 10 H 14 FeN2O8, C6H2FeN3O7 + , C2H2Fe, C 10 H6FeN2, C6H 15 FeN3O7, C 72 H 124 FeO8 -2 , FeH 22 N2O 15 S2, C 40 H 78 FeO4, FeH2N2O6 +2 , C 44 H 86 FeO4, C 10 H 20 FeN2O8S2, C 20 H 38 FeO4, C 36 H 66 FeO4, C 24 H 46 FeO4, C 29 H 26 FeP + , C 36 H64 FeO6, C 14 H 26 FeO4, C 26 H 28 FeNP, C 28 H 54 FeO4, C 36 H 32 FeN4O4, C 36 H 36 FeN4O8, C6H9FeNO7 + , C5H6FeO2, C4H 11 BFeO4, C8H 19 BFeO4, C4H4FeO4S2, C6H6FeO7, C 18 H 34 FeO4, C 12 H 20 FeO 13 , C4H4FeO6, C5H7FeNO3, Fe3H8O4, C2FeN2S2, FeH2O2, Fe3H2O4, C 44 H 28 FeN4, C2H6FeO5, Fe2H6O 11 S2, C3H4FeN2O3, Fe3H2O9P2, C6H 14 Fe3N3O7 -3 , C4H 10 FeN2O6, Cl2FeH2O, FeO4W, C6H5FeO3P, C6H8FeO7, FeTe, C4H2FeO4, C 20 H 20 Cl2FeN8, C 14 H 12 FeO6, C3H3FeO7P, C4H7FeNO4, FeO3Si, Cl2FeH 12 O6, Cl2FeH2O9, FeH 10 O9S, FeH 12 O 10 S, C8H 17 FeO3P, C4H 14 FeO8, Fe3H 16 O 16 P2, F6FeSi, C 72 H 42 FeN6Na6O 22 S7, FeH4O5S, C 39 H 30 FeN6O4S, C 40 H 50 O4, C4H10 FeN2O4, C2H4FeN2O4S, Br2FeH2O, C 98 H 200 FeN 10 、C 36 H 21 FeN9O 10 S、C 10 H 10 Fe, C2H6FeN2, F6FeH 12 O6Si、C 48 H 48 FeN6O4S、FeO4S、C2H 10 FeN2O8S2、C 44 H 27 FeN5O、C 30 H 24 FeN6O4S、C6H8O6、C6H7NaO6、FeH4O2 +2 、FeH2O +2 、C3H7FeNO7S、C 30 H 18 FeN3NaO6、C2H 18 FeN2O 12 S2、C4H4FeO4、C7H7FeN4O + 、Br2Fe、C 18 H 22 Cl2FeN2、C 32 H 28 FeN6O6S2、C 12 H 14 MgO 12 、C2H5FeNO6S、C 45 H 60 FeN2O8、C 30 H 22 Cl2FeN2、C 38 H 26 FeN8O2S2、C 30 H 28 FeN2O6、C 14 H 12 Cl6FeO4、C 12 H 14 Fe、C 36 H 36 Cl2FeN6O8、C 17 H 14 FeN4O4S、C 24 H 30 FeN4O4、C 34 H 32 ClFeN4O6、C12 H 12 Fe, Fe3H 14 O 12 P2 +6 , C 32 H 16 FeN8, FeS2, C 16 H 15 FeNO2 +2 , C 29 H 20 FeO6, C 23 H 28 FeO2, C 11 H 10 FeO2, C 13 H 14 FeO2, C 12 H 12 FeO2, C 46 H 48 FeN4O6 +2 , C 47 H 59 FeN 13 O8 +2 , C 46 H 59 FeN 13 O8 +2 , C 48 H 62 FeN 12 O8S +2 , C 50 H 65 FeN 13 O8 +2 , C 48 H 63 FeN 13 O8 +2 , C 48 H 62 FeN 12 O8S +2 , C 55 H 76 FeN 14 O9 +2 , C 25 H 19 FeN3, C 15 H 17 FeN3OS +2 , C 22 H 23 FeN3OS +2 , C 26 H 28 ClFeN3, C 28 H 33 ClFeN4, C27 H 31 ClFeN4, C 29 H 35 ClFeN4, C 30 H 37 ClFeN4, C 28 H 33 ClFeN4, C 27 H 30 ClFeN3, C 26 H 28 ClFeN3, C 29 H 35 ClFeN4, C 27 H 30 ClFeN5O +2 , C 41 H 38 ClFeN5O3 +2 , C 42 H 41 FeN5O3 +2 , C 41 H 38 FFeN5O3 +2 , C 42 H 47 FeN5O3 +2 , C 43 H 49 FeN5O3 +2 , C 42 H 41 FeN5O3 +2 , C 42 H 40 ClFeN5O3 +2 , C 42 H 40 ClFeN5O3 +2 , C 42 H 40 FFeN5O3 +2 , C 41 H 45 FeN5O3 +2 , C 42 H 47 FeN5O3 +2 , C 41 H 39 FeN5O3 +2 , C 22 H 25 FeN5O5 +2 , C 24 H 23 ClFeN4O2 +2 , C 24 H 23FFeN4O2 +2 、C 24 H 24 FeN4O2 +2 、C 15 H 21 FeN3S +2 、C 29 H 34 FeN4O2 +2 、C 28 H 31 ClFeN4O2 +2 、C 28 H 31 FFeN4O2 +2 、C 30 H 35 ClFeN4O3 +2 、C 30 H 35 FFeN4O3 +2 、C 28 H 32 FeN4O2 +2 、C 27 H 30 FeN4O2 +2 、C 26 H 27 ClFeN4O2 +2 、C 30 H 36 FeN4O3 +2 、C 28 H 31 ClFeN4O3 +2 、C 28 H 31 FFeN4O3 +2 、C 28 H 32 FeN4O3 +2 、C 27 H 29 ClFeN4O3 +2 、C 26 H 27 FFeN4O2 +2 、C 26 H 28 FeN4O2 +2 、C 26 H 28 FeN4O2 +2 、C 27 H 29 FFeN4O3 +2 、C 27 H 30 FeN4O3 +2 、C26 H 27 ClFeN4O3 +2 、C 26 H 27 FFeN4O3 +2 、C 26 H 28 FeN4O3 +2 、C 25 H 25 ClFeN4O3 +2 、C 25 H 25 FFeN4O3 +2 、C 25 H 26 FeN4O3 +2 、C 24 H 23 ClFeN4O3 +2 、C 24 H 23 FFeN4O3 +2 、C 24 H 24 FeN4O3 +2 、C 25 H 25 ClFeN4O2 +2 、C 25 H 25 FFeN4O2 +2 、C 25 H 26 FeN4O2 +2 、C 25 H 26 FeN4O2 +2 、C 29 H 32 ClFeN7 +2 、C 33 H 32 ClFeN7 +2 、C 22 H 27 ClFeN3RuS + 、C 18 H 19 ClFeN3RuS + 、C 19 H 19 BFeO3 +2 、C 28 H 25 ClFeN4O +2 、C 31 H 38 FeN4O3、C 29 H 34 FeN4O3、C31 H 41 FeN3O, C 28 H 32 FeN4O3, C 26 H 29 FeN3O2, C 26 H 30 FeN2O, C 31 H 36 FeN4O3, C 30 H 35 FeN5O4, C 29 H 35 FeN5O3, C 32 H 41 FeN5O3, C 35 H 38 FeN4O3, C 32 H 40 FeN4O3, C 19 H 13 BBr2F2FeO2, C 19 H 14 BClF2FeO2, C 19 H 14 BBrF2FeO2, C 19 H 15 BF2FeO2, C 21 H 20 FeO4, C 20 H 18 FeO3, C 20 H 18 FeO3, C 20 H 18 FeO3, C 19 H 14 F2FeO2, C 19 H 14 Br2FeO2, C 19 H 15 BrFeO2, C 14 H 12 FeO3, C 21 H 19 BF2FeO4, C 20 H 17 BF2FeO3, C 20 H 17 BF2FeO3, C 20 H 17 BF2FeO3, C 19 H 13 BF4FeO2, C 19 H 13 BCl2F2FeO2, C 21 H29 AuCl2FeN4S + 、C 30 H 24 Cl2FeN6 +2 、C 22 H 21 Cl2FeN3 +2 、C 23 H 22 FeN6 +2 、C 21 H 19 FeN7 +2 、C 23 H 24 FeN6O +2 、C 47 H 64 FeN 14 O9、C 46 H 60 FeN 12 O 10 、C 41 H 53 FeN 11 O7、C 47 H 65 FeN 15 O8、C 45 H 59 FeN 13 O9、C 42 H 54 FeN 12 O7、C 43 H 67 FeN 15 O8、C 48 H 65 FeN 13 O8、C 47 H 64 FeN 12 O8、C 54 H 77 FeN 17 O9、C 51 H 71 FeN 15 O 10 、C 19 H 16 FeO2、C 44 H 48 FeN9O 17 P3、C 13 H9Cl2FeN3O6S、C 19 H 15 FeNO3、C 20 H 18 FeO2、C 20H 18 FeO3, C 21 H 20 FeO3, C 17 H 20 FeN2O2, C 18 H 15 FeNO, C 17 H 14 FeOS, C 17 H 14 FeOS, C 17 H 14 FeO2, C 22 H 22 FeO4, C 20 H 18 FeO2, C 20 H 18 FeO2, C 19 H 14 Cl2FeO, C 21 H 20 FeO3, C 48 H 28 FeN4O8, C 17 H 15 FeNS, C 34 H 30 FeN4O4 -2 , C 30 H 26 Br2FeN4O4, C 10 H 18 FeN2O7 +2 , C 14 H 12 FeO4, C 44 H 20 Cl8FeN4, C 64 H 64 FeN8O 12 S4, C 56 H 56 FeN8O8S4, C 56 H 44 Br8FeN4, C 56 H 52 FeN4, C 52 H 40 FeN8O 12 S4, C 44 H 32 FeN8O8S4, or C 44 H 28 FeN4. In some other cases, the source of iron has the chemical formula C a H b Fe c Od N e S f Br g Cl h P i Na j As k K l Al m C rn V o I p B q F r Te s W t, where the coefficients a, b, c, d, e, f, g, h, i, j, kl, m, n, o, p, q, r, s, t can be equal to 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or any integer between 21 and 1000000000000, where the C, H, Fe, O, N, S, Br, Cl, P, Na, As, K, Al, Cr, V, I, B, F, Te and W atoms preferably occupy the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, eighteenth, nineteenth, twentieth positions respectively. In some cases, at least one atom of the chemical formula can occupy any one of the 20 positions of the formula.In some other cases, the source of iron may include chemical functional groups selected from the group consisting of: alkanes (R(CH2)nH), alkenes (R2C═CR2), alkynes (RC≡CR'), benzene derivatives (RC6H5); groups containing halogens, haloalkanes (RX), oxygen-containing, alcohol groups (ROH), carbonyls (RCOR'), aldehydes (RCHO), acyl halides (RCOX), carbonates (ROCOOR'), carboxylates (RCOO-), carboxylic acids (RCOOH), esters (RCOOR'), methoxy groups (ROCH3), hydroperoxides (ROOH), peroxides (ROOR'), ethers (ROR'), hemiacetals (RCH(OR')(OH)), hemiketals ((RC(OR″)(OH)R'), acetals (RCH(OR')(OR″)), ketals ((RC(OR″)(OR″′)R'), ortho esters (RC(OR')(OR″)(OR″′)), heterocycles (PhOCOPh), orthocarbonates (C(OR)(OR')(OR″)(OR″)), nitrogen-containing, amides (RCONR2), amines (RNH2, R2NH, R3N, R4N+), imines ((RC(═NH)R', RC(═NR”)R’, RC(═NH)H, RC(═NR')H, acylimides ((RCO)2NR'), azides (RN3), azo compounds (RN2R'), cyanates (ROCN, RNCO), nitrates (RONO2), nitriles (RCN, RNC), nitrites (RONO), nitro compounds (RNO2), nitroso compounds (RNO), oximes (RCH═NOH), pyridine derivatives (RC5H4N), sulfur-containing groups, thiols (RSH), sulfides (RSR'), disulfides (RSSR'), sulfoxides (RSOR'), sulfones (RSO2R'), sulfinic acids (RSO2H), sulfonic acids (RSO3H), thiocyanates (RSCN, RNCS), thioketones (RCSR'), thials (RCSH), phosphorus-containing groups, phosphines (R3P), phosphonic acids (RP(═O)(OH)2), phosphates (ROP(═O)(OH)2), phosphodiesters (HOPO(OR)2), boron-containing groups, boric acids (RB(OH)2), borate esters (RB(OR)2), boric anhydrides (R2BOH), borate anhydrides (R2BOR), and several combinations of these groups. In some other cases, the iron source may be an iron chelator. In some cases, the amount of the iron source is the amount or concentration of the iron source or iron, preferably derived from the iron source, preferably from the pre-growth and / or growth medium.

[0588] The present invention also relates to a method according to the present invention, wherein the iron or iron source in the pre-growth medium consists of Fe 2+ and / or Fe 3+ or comprises Fe 2+ and / or Fe3+ .

[0589] In some cases, when the iron source contains Fe2 in its chemical formula, the iron source contains Fe 2+ . In some other cases, when the iron source contains Fe 3 in its chemical formula, the iron source contains Fe 3+ .

[0590] The present invention relates to a method according to the present invention, wherein the concentration of iron or an iron source in the pre-growth medium is less than 20 μM. In some cases, the concentration of iron or an iron source in the pre-growth medium is less than 10 100 、10 50 、10 20 、10 10 、10 5 、10 3 、10 2 or 20 μM. In some other cases, the concentration of iron or an iron source in the pre-growth medium is greater than 0, 10 -50 、10 -20 、10 -5 、10 -1 、1, 5, 10 or 20 μM. In some other cases, the concentration of iron or an iron source in the pre-growth medium is between 10 -50 and 10 50 、10 -10 and 10 10 、10 -10 and 10 5 、10 -10 and 10 3 or between 10 -10 and 1 μM.

[0591] The present invention relates to a method according to the present invention, wherein the growth medium contains iron or at least one iron source, and the nature and / or amount of the iron or iron source allows the generation of nanoparticles by cells capable of generating nanoparticles and cell growth.

[0592] The present invention relates to a method according to the present invention, wherein the iron source of the growth medium is the same as the iron source of the pre-growth medium.

[0593] The present invention relates to a method according to the present invention, wherein the concentration of iron or an iron source in the growth medium is greater than or equal to the concentration of iron or an iron source in the pre-growth medium.

[0594] In some cases, the concentration of iron or an iron source in the growth medium is less than 10 100 、10 50 、10 20 、10 10 、10 5 、10 3 、102 or 20 μM. In some other cases, the concentration of iron or an iron source in the growth medium is greater than 0, 10 -50 , 10 -20 , 10 -5 , 10 -1 , 1, 5, 10 or 20 μM. In some other cases, the concentration of iron or an iron source in the growth medium is between 10 -50 and 10 50 , 10 -10 and 10 10 , 10 -10 and 10 5 , 10 -10 and 10 3 and between 10 -10 and 1 μM.

[0595] The present invention relates to a method according to the present invention, wherein the growth medium is supplemented with a fed-batch medium.

[0596] In one embodiment of the present invention, the fed-batch medium contains at least one source common to the pre-growth and / or growth medium. In some cases, the concentration of this source is equal to or at least 1.00001, 1.1, 2, 5, 10, 10 3 or 10 5 in the fed-batch medium than in the pre-growth and / or growth medium / media. In some other cases, the concentration of this source is at least 10 5 , 10 3 , 10, 1, 1.1 or 1.00000001 lower in the fed-batch medium than in the pre-growth and / or growth medium / media.

[0597] The present invention relates to a method according to the present invention, wherein the fed-batch medium contains a concentration greater than 10 -50 , 10 -20 , 10 -10 , 10 -5 , 10 -1 , 1, 5, 10, 20, 50, 100, 10 3 or 10 5 μM. In some other cases, the fed-batch medium contains a concentration lower than 10 50 , 10 20 , 10 10 , 10 5 , 10 2 , 10 -2 , 10 -10 or 10 -20 M of iron or an iron source. In some other cases, the fed-batch medium contains iron or an iron source, the concentration of which is between 10 -50and 10 50 , 10 -15 and 10 15 , 10 -10 and 10 5 , 10 -5 and 10 5 , 10 -3 and 10 3 between 10 and 10 μM, or between 0.5 nM and 50 μM, preferably before adding the fed-batch medium to the growth medium.

[0598] In one embodiment of the present invention, the fed-batch medium is acidic or has a pH below 7, 6, 5, 4 or 3, preferably below 2. In some cases, the fed-batch medium has a pH greater than 0 or 1.

[0599] The present invention relates to a method according to the present invention, wherein the fed-batch medium is added to the growth medium at a rate between 10 -15 liters per hour and 10 15 liters per hour or between 10 -15 micromoles of iron per hour and 10 15 micromoles of iron per hour. In some cases, the fed-batch medium is added to the growth medium at a low rate, preferably at a rate lower than 10 100 , 10 50 , 10 20 , 10 10 , 10 5 , 10 2 , 10, 5, 1, 10 -2 , 10 -3 , 10 -5 or 10 -10 liters of fed-batch medium per minute or micromoles of iron per minute, preferably when the number of cell divisions in the growth medium is low, preferably lower than 10 20 , 10 10 , 10 5 , 10 3 , 10 cell divisions per second or per hour or per day or per month. In some other cases, the fed-batch medium is added to the growth medium at a high rate, preferably at a rate greater than 10 -100 , 10 -50 , 10 -20 , 10 -10 , 10 -5 , 10 -2 , 1, 5, 10, 10 2 , 10 3 , 10 5 or 10 10Liters of fed-batch medium per minute or micromoles of iron per minute, preferably when the number of cell divisions in the growth medium is large, preferably greater than 1, 2, 5, 10, 10 3 、10 5 、10 10 or 10 20 , cell divisions per second or per hour or per day or per month. In some cases, between two sub-steps of the growth step, the flow rate of the fed-batch medium is reduced, preferably by a factor greater than 1.0000001, 1.00001, 1.0001, 1.01, 1.1, 1.2, 1.5, 2, 5, 10, 10 3 、10 5 、10 7 or 10 9 . In some other cases, between two sub-steps of the growth step, the flow rate of the fed-batch medium is increased, preferably by a factor greater than 0000001, 1.00001, 1.0001, 1.01, 1.1, 1.2, 1.5, 2, 5, 10, 10 3 、10 5 、10 7 or 10 9 .

[0600] In one embodiment of the present invention, before adding the growth medium, the iron concentration of the fed-batch medium is at least 1.000001, 1.001, 1, 1.5, 2, 5, 10, 10 2 、10 3 or 10 5 times greater than the iron concentration of the growth medium.

[0601] In another embodiment of the present invention, preferably after adding the growth medium, the fed-batch medium becomes part of the growth medium.

[0602] The present invention relates to a method according to the present invention, wherein the pre-growth and / or growth medium according to the present invention contains only one vitamin selected from calcium pantothenate, folic acid, inositol, nicotinic acid, p-aminobenzoic acid, pyridoxine hydrochloride, riboflavin, thiamine hydrochloride and any derivatives of these vitamins.

[0603] In some cases, the pre-growth and / or growth medium may contain less than 10 50 、10 20 、10 10 、10 5 、10 3 、100, 75, 50, 25, 10, 5, 3, 2 or 1 vitamin or different vitamins. In some cases, the different vitamins may be those containing at least 1, 2, 5, 10, 10 2 、103 , 10 5 , 10 10 , 10 20 or 10 50 kinds of vitamins of different chemical elements. In some other cases, the pre-growth and / or growth medium contains more than 1, 2, 5, 10, 10 2 , 10 3 , 10 5 or 10 10 kinds of vitamins or different vitamins.

[0604] In certain cases, the vitamin can be a water-soluble vitamin. In some other cases, the vitamin can be a fat-soluble vitamin. In some other cases, the vitamin can belong to vitamin A, D, E, K, B1, B2, B3, B5, B6, B7, B9, B12 or vitamin C. In some other cases, the vitamin is not produced by the cells that generate the nanoparticles. In some other cases, the vitamin can be a vitamin for treating diseases caused by or related to vitamin deficiency.

[0605] In another embodiment of the present invention, the vitamins are selected from: adenosylcobalamin, aminobenzoic acid, ascorbic acid, biotin, calcium D-(+)-pantothenate, carotene thiamine, carotenoid beta, cholecalciferol (D3), cyanocobalamin, cyanocobalamin, ergocalciferol (D2), folic acid, folic acid, folinic acid, hydroxocobalamin, inositol, menaquinone (K2), methylcobalamin, niacin, nicotinamide, nicotinamide, nicotinamide riboside, niacin, pantothenic acid, phylloquinone (K1), pyridoxal, pyridoxamine, pyridoxol, lipoic acid, pyridoxal, pyridoxamine, pyridoxol, pyridoxine hydrochloride, retinal, retinoic acid, retinol, riboflavin, thiamine, tocopherol or tocotrienol, and derivatives or combinations of one or several of these vitamins.

[0606] In another embodiment of the present invention, at least one vitamin contained in the growth medium is biotin, folic acid, riboflavin, niacin or thiamine.

[0607] In some cases, the vitamins contained in the pre-growth medium are the same as those contained in the growth medium. In some other cases, the vitamins contained in the pre-growth medium are different from those contained in the growth medium.

[0608] The present invention also relates to the method according to the present invention, wherein the concentration of at least one vitamin contained in the pre-growth and / or growth medium / media is lower than 10 100 , 10 50 , 10 20 , 10 10 , 10 5 , 103 , 10, 1, 10 -1 , 10 -3 , 10 -4 , 10 -6 , 10 -9 , 10 -20 , 10 -50 or 10 -100 M, or preferably less than 0.002 mol / L.

[0609] In another embodiment of the present invention, the pre-growth and / or growth medium / medium comprises at least one vitamin or one chemical element wherein the concentration of at least one vitamin is greater than 10 -100 , 10 -50 , 10 -20 , 10 -10 , 10 -5 , 10 -4 , 10 -3 , or 10 -1 , 1, 10, 10 2 , 10 3 , 10 5 or 10 10 M.

[0610] The present invention also relates to a method according to the invention, wherein the pre-growth and / or growth medium comprises at least one vitamin in a lower concentration than a source of carbon, nitrogen, sulfur, sulfate, phosphorus, phosphate, calcium, potassium, magnesium, oxygen, hydrogen and / or iron, preferably at least 1.0001, 1.2, 1.5, 2, 5, 10, 10 3 or 10 5 In some cases, cells do not need large amounts of the vitamin to grow, divide and / or synthesize nanoparticles.

[0611] The present invention also relates to a method according to the invention, wherein the pre-growth and / or growth medium / medium comprises, per gram or per milliliter of growth or pre-growth medium, less than: i) 1 mg yeast extract, ii) 1 mg at least one component of yeast extract, iii) 1 mg peptone, iv) 1 mg at least one peptone component, v) 1 mg CMR reagent, vi) 1 mg at least one chelating agent, vii) 1 mg at least one amino acid, viii) 1 mg toxic or cytotoxic compound, and / or ix) 1 mg at least one heavy metal.

[0612] In one embodiment of the invention, the pre-growth and / or growth medium comprises yeast extract, peptone, CMR reagent, chelating agent, amino acid, toxic or cytotoxic compound and / or heavy metal in a concentration of less than 10 100 , 10 50 , 1020 、 10 10 、 10 5 、 10 2 、 10, 1, 10 -3 or 0 -5 μg of yeast extract, peptone, CMR agent, chelating agent, amino acids and / or heavy metals per liter or per milliliter of pre - growth and / or growth medium / media. This occurs in some cases when the yeast extract, peptone, CMR agent, chelating agent, amino acids, toxic or cytotoxic compounds and / or heavy metals have been removed from the pre - growth and / or growth medium or are not included in the pre - growth..

[0613] In one embodiment of the present invention, the first and / or second medium / media contains yeast extract, peptone, amino acids and / or heavy metals in a concentration greater than 10 -100 、 10 -50 、 10 -20 、 10 -10 、 10 -5 、 10 -2 、 10 -1 、 1, 10, 10 3 or 10 5 μg of yeast extract, peptone, amino acids and / or heavy metals per liter or per milliliter of pre - growth and / or growth medium / media. This may occur in some cases when the yeast extract, peptone, CMR agent, chelating agent, amino acids and / or heavy metals have been (preferably not deliberately) added to the pre - growth and / or growth medium / media.

[0614] In one embodiment of the present invention, the yeast extract is or contains peptides, amino acids, purine bases, pyrimidine bases and / or water - soluble vitamins of group B. In one embodiment of the present invention, the amino acids are alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and / or valine.

[0615] In one embodiment of the present invention, the heavy metals are arsenic (As), cadmium (Cd), chromium (Cr), copper (Cu), mercury (Hg), nickel (Ni), lead (Pb), selenium (Se) and / or zinc (Zn).

[0616] In one embodiment of the present invention, the reagent having carcinogenic, mutagenic or toxic properties for reproduction (also called CMR reagent) is preferably nitrilotriacetic acid, trisodium salt and / or boric acid.

[0617] In one embodiment of the present invention, a toxic or cytotoxic compound is a compound that causes cell death, preferably a cell that generates nanoparticles, preferably when introduced into a pre-growth and / or growth medium, preferably at a concentration greater than 10 -10 、10 -5 、10 -2 、1、5、10、10 3 or 10 μM, preferably at a concentration between 10 -10 and 10 10 μM.

[0618] In one embodiment of the present invention, the pre-growth and / or growth medium / culture medium contains / does not contain minerals selected from the group consisting of: selected from: C6H6NO6Na3, trisodium nitrilotriacetate, MnO4SH2O, manganese(II) sulfate monohydrate, NaCl, sodium chloride, CoN2O6 6H 2O, cobalt(II) nitrate hexahydrate, O 4SZn 7H 2O, zinc sulfate heptahydrate, CuO 4S 5H2O, copper(II) sulfate pentahydrate, AlKO 8S2 12H 2O, potassium alum, H3BO3, boric acid, Na2MoO4·2H2O, sodium molybdate dihydrate, Cl2Ni6H2O, nickel(II) chloride hexahydrate, Na2SeO3, sodium selenite, and derivatives or combinations of one or more of these compounds.

[0619] In one embodiment of the present invention, the pre-growth and / or growth medium contains a certain concentration of minerals that is less than 10 100 、10 50 、10 20 、10 10 、10 5 、10 2 、10、1、10 -1 、10 -3 or 10 -5 μg of minerals per liter or milliliter of growth medium. In some cases, this may occur when the minerals are removed from the pre-growth and / or growth medium.

[0620] In one embodiment of the present invention, the first (pre-growth) and / or second growth medium contains a certain concentration of minerals that is greater than 10 -100 、10 -50 、10 -20 、10 -10 、10 -5 、10 -2 、10 -1 、1、10、10 3 or 10 5μg mineral per litre or millilitre of growth medium. In some cases this may occur when minerals have been (preferably unintentionally) removed from the growth and / or growth medium / culture medium.

[0621] The invention also relates to a method according to the invention comprising an additional step of purifying the high-purity iron oxide nanoparticles by removing at least one impurity from the nanoparticles.

[0622] In one embodiment of the present invention, an additional step of purifying the high purity iron oxide nanoparticles consists in removing impurities from the nanoparticles and / or denaturing and / or destroying impurities contained in the nanoparticles.

[0623] In one embodiment of the invention, the additional step of purifying the nanoparticles is preceded by the aforementioned step of isolating or extracting the nanoparticles from the cells. In some cases, the aforementioned step is a step of recovering the nanoparticles. In some cases, the aforementioned step is performed by i) mixing the cells, preferably obtained from the growing step, with a detergent such as KOH or NaOH, ii) purifying the nanoparticles at a temperature of greater than -270, -250, -200, -150, -100, -50, -30, -10, -5, 0, 5, 10, 20, 30, 50, 75, 100, 150, 200, 500, 10 3 , 10 5 or 10 10 ℃, or between -270 and 10 10 Between - 100 and 10 5 or heating the cells between 0 and 100°C, iii) with a temperature gradient greater than 10 -50 , 10 -20 , 10 -10 , 10 -5 , 10 -3 , 10 -1 , 1, 2, 5, 10, 10 3 , 10 5 or 10 10 °C / hour, minute or second, or between 10 -50 and 0 10 ℃ per hour, minute or second, iii) applying pressure to the cells, preferably greater than 100, 500, 10 3 , 10 4 , 10 5 , 10 6 , 10 7 , 10 8 or 10 9 atmospheres, or between 1 and 10 9 atmospheres, for example using French pressure, and / or, iv) preferably at a pressure greater than 10 -50, 10 -20 , 10 -5 , 10 -1 , 1, 10, 10 2 , 10 3 , 10 5 or 10 10 Ultrasonically treat the cells with the power of W.

[0624] In another embodiment of the present invention, an additional step of purifying the nanoparticles can remove: i) impurities of greater than 10 -20 , 10 -10 , 10 -5 , 10 -2 , 10 -1 , 1, 5, 10, 25, 50, 75, 80 or 90% by mass of impurities ii) higher than 10 -50 , 10 -20 , 10 -10 , 10 -5 , 10 -1 , 1, 5, 10, 10 3 , 10 5 or 10 10 μg of impurities per gram of nanoparticles.

[0625] In one embodiment of the present invention, the impurities preferably removed by an additional step of purifying the nanoparticles are carbon or carbon-containing materials and / or not iron oxide. Preferably, such / these impurities are located in the coating of the nanoparticles.

[0626] In one embodiment of the present invention, impurities are removed from the core and / or coating of the nanoparticles, preferably from the coating of the nanoparticles. Preferably, the removed impurities are shallow impurities. In some other cases, the removed impurities are deep impurities.

[0627] The present invention also relates to an additional step of a method for purifying high-purity iron oxide nanoparticles by removing at least one impurity from the nanoparticles, including at least one heating step, wherein the temperature of the nanoparticles is increased to temperature T0 and then maintained at T0 during the heating time, and the heating time preferably includes between 1 second and 1 minute, 1 second and 1 hour, 1 second and 12 hours, 1 second and 1 day, 1 second and 1 week, 1 second and 1 month, or 1 second to 1 year, wherein T0 preferably includes between -200 and 10 5 , -100 and 10 5 , -50 and 10 5 , -10 and 10 5 , 0 and 10 5 , 10 and 10 5 , 20 and 10 5 , 30 and 10 5 , 100 and 105 , 200 and 10 5 , 100 and 10 4 , 100 and 10 3 , or between 100 and 500 °C.

[0628] The present invention also relates to an additional step of a method for purifying high-purity iron oxide nanoparticles by removing at least one impurity from the nanoparticles, which comprises at least two heating steps, wherein:

[0629] - During step 1, the temperature of the nanoparticles is raised to temperature T1 and then maintained at T1 for a heating time between 1 second and 20 years, wherein T1 is between 150 °C and 250 °C.

[0630] - During step 2, the temperature of the nanoparticles is raised to temperature T2 and then maintained at T2 for a heating time between 1 second and 20 years, wherein T2 is between 350 °C and 450 °C.

[0631] In some cases, the method for purifying high-purity iron oxide nanoparticles or the additional step of the heating step may be referred to as a purification method.

[0632] The present invention relates to a method for removing at least one impurity from high-purity iron oxide nanoparticles, including an additional step between step 1 and step 2, wherein the temperature of the nanoparticles is raised to temperature T3 and then maintained at T3 for a heating time between 1 second and 20 years. Wherein T3 is 250 °C to 350 °C.

[0633] In some cases, during a heating time of less than 100 years, 50 years, 20 years, 10 years, 5 years, 2 years, 1 year, 11 months, 6 months, 3 months, 2 months, 1 month, 3 weeks, 2 weeks, 1 week, 6 days, 5 days, 3 days, 1 day, 23 hours, 12 hours, 6 hours, 1 hour, 50 minutes, 30 minutes, 20 minutes, 10 minutes, 5 minutes, 2 minutes, 1 minute, 50 seconds, 30 seconds, 10 seconds, 1 second, 1 millisecond or 1 microsecond, the temperature of the nanoparticles remains at temperature T1, T2 and / or T3. In some other cases, during a heating time of greater than 1 microsecond, 1 millisecond, 1 second, 10 seconds, 30 seconds, 50 seconds, 1 minute, 2 minutes, 5 minutes, 10 minutes, 1 hour, 6 hours, 12 hours, 23 hours, 1 day, 3 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 6 months, 11 months, 1 year, 2 years, 5 years, 10 years, 20 years, 50 years or 100 years, the temperature of the nanoparticles remains at temperature T1, T2 and / or T3. In some other cases, during a heating time between 1 microsecond and 100 years, 1 second and 20 years, 1 second and 1 year, 1 second and 1 month, 1 second and 1 day, 1 minute and 1 day, 5 minutes and 1 day, 10 minutes and 12 hours, 30 minutes and 6 hours, or 30 minutes and 3 hours, the temperature of the nanoparticles remains at temperature T1, T2 and / or T3.

[0634] In one embodiment of the present invention, the heating time is greater than the duration for the temperature to rise to T1, T2 and / or T3, preferably greater than 1.001, 1.1, 1.5, 2, 5, 10, 10 3 , 10 5 or 10 10 times.

[0635] In certain cases, T1 is between -273 °C and 250 °C, -200 °C and 250 °C, -100 °C and 250 °C, 0 and 250 °C, 50 °C and 250 °C, 150 °C and 250 °C, or between 180 °C and 220 °C. In some other cases, T2 includes between 200 and 10 5 , 250 and 10 5 , 300 and 10 5 , 350 and 10 5 , 350 and 10 3 , 350 and 500, 350 and 450, or between 360 and 400 °C. In certain cases, T3 includes between -273 and 10 5 , -200 and 10 3 , -100 and 500, -50 and 200, 0 and 500, 100 and 500, 200 and 500, 200 and 400, or between 250 and 350 °C.

[0636] In yet another embodiment of the present invention, T3 is included between T1 and T2. In some cases, T3 is lower than T2, preferably higher than 1.0001, 1.1, 1.2, 1.5, 2, 5, 10, 10 3 or 10 5 times. In some other cases, T3 is greater than T1, preferably greater than higher than 1.0001, 1.1, 1.2, 1.5, 2, 5, 10, 10 3 or 10 5 times.

[0637] In one embodiment of the present invention, the temperature of the nanoparticles is the temperature for heating the nanoparticles and / or the heating device or furnace containing the nanoparticles, preferably before, during, or after treating the nanoparticles by a purification method.

[0638] In one embodiment of the present invention, the temperature interval separating T1 and T2, designated as [T1, T2], is such that: i), the nanoparticles exhibit a maximum change or loss in weight or mass as a function of temperature, and / or ii) the derivative of the change or loss in weight or mass of the nanoparticles as a function of time is maximum.

[0639] In one embodiment of the present invention, the ratio [%W(T2)-%W(T1)] / (T2-T1), where %W(T2) and %W(T1) are the weight or mass percentages of the nanoparticles at T2 and T1 respectively, 10 -50 、10 -30 、10 -20 、10 -10 、10 -5 、0.01, 0.02, 0.03, 0.05, 0.1, 0.2, 0.3, 0.5, 1, 10 or 10 5 % / °C. In some cases, when the proportion of the carbon mass percentage in the nanoparticles (preferably before treating the nanoparticles by a purification method) is large, this ratio is large, preferably greater than 10 -20 、10 -10 、10 -5 、10 -2 、10 -1 、1, 5, 10, 20, 50, 75, 85, 95 or 100% / °C.

[0640] In another embodiment of the present invention, the ratio [%W(T2)-%W(T1)] / (T2-T1) is lower than 10 50 、10 30 、10 20 、10 10 、10 5, 10, 5, 2, 1, 0.5, 0.05, 10 -3 , 10 -5 , 10 -10 or 10 -20 % / °C. In some cases, when the proportion of the carbon mass percentage in the nanoparticles (preferably before or after treating the nanoparticles by a purification method) is very low, this ratio is very low, preferably less than 100, 95, 80, 70, 50, 30, 20, 10, 5, 2, 1, 10 -1 , 10 -3 , 10 -5 , 10 -10 or 10 -20 % / °C.

[0641] In one embodiment of the present invention, the temperature of the nanoparticles is maintained at T1, T2 and / or T, when the change of T1, T2 and / or T3 is less than 10 5 , 10 3 , 90, 80, 70, 60, 50, 40, 30, 20, 10, 5, 3, 2, 1, 10 -5 , 10 -10 or 10 -20 %. In certain cases, for each temperature T1, T2, and / or T3, this percentage is equal to T maxi -T mini / T avi , where T maxi , T mini , and T avi (i = 1, 2, 3) is the maximum value, minimum value, and average temperature reached during the heating time or during the heating step, preferably after the temperature is maintained at the temperature T1, T2, and / or T3 or when the temperature is maintained at the temperature T1, T2, and / or T3. In some cases, when the furnace or heating device can maintain the temperature stable without large fluctuations and / or when the nanoparticles are not prone to endothermic and / or exothermic reactions, this percentage is low. In some cases, an endothermic reaction is a reaction in which heat or energy is transferred from the medium around the nanoparticles to the nanoparticles. In some other cases, an exothermic reaction is a reaction in which heat or energy is transferred from the nanoparticles to the medium around the nanoparticles.

[0642] In one embodiment of the present invention, when the temperature change of the nanoparticles is higher than 10 5 , 10 3 , 90, 80, 70, 60, 50, 40, 30, 20, 10, 5, 3, 2, 1, 10 -5 , 10 -10 or 10 -20When it is [X]%, the temperature of the nanoparticles does not remain at T1, T2, and / or T3. In some cases, this percentage is large when the furnace or heating device cannot maintain temperature stability without large fluctuations and / or when the nanoparticles are prone to endothermic and / or exothermic reactions.

[0643] In one embodiment of the present invention, the temperature T1 and / or T2 is determined by the following formula:

[0644] i), measure the change in the weight or mass percentage of the nanoparticles with temperature when the nanoparticles are heated between two temperatures T T<T1 and T T>T2 where T T<T1 is lower than T1 and T T> T2 is greater than T2,

[0645] ii) measure or represent or consider or examine or use at least one peak of the derivative of this percentage change as a function of temperature,

[0646] iii) estimate or infer the temperature range where the change in the weight or mass percentage of the nanoparticles with temperature is the largest, where the minimum and maximum temperatures of this temperature range are T1 and T2 respectively,

[0647] iv) estimate or derive two temperatures T1 and T2 from the positions of at least one peak in the derivative curve of the change in the weight or mass percentage of the nanoparticles, obtaining the minimum values at the start and end of the derivative curve peaks respectively, T3 is obtained from the maximum value in the middle of the derivative curve peaks, and

[0648] v) estimate or infer the temperatures T1, T2, T3 from the positions of at least one peak in the curve of the heat flux of the nanoparticles with temperature, preferably located at the start of at least one peak.

[0649] In some cases, the temperature where the derivative of the change in the weight or mass percentage of the nanoparticles starts to show a decrease with temperature is at the start of the peak. In some other cases, the temperature where the derivative of the change in the weight or mass percentage of the nanoparticles starts to show an increase with temperature is at the end of the peak.

[0650] In another embodiment of the present invention, the heat flux of the nanoparticles is the heat flux generated by the nanoparticles or released from the nanoparticles or originating from the nanoparticles, preferably when the nanoparticles are heated with a heating device such as a furnace. Preferably, the heat flux can be measured with a device or using a thermal analysis method, or using differential thermal analysis (DTA) or using differential scanning calorimetry (DSC).

[0651] In one embodiment of the present invention, heating step i of the purification method according to the present invention, where i is preferably an integer greater than or equal to 0, includes at least one of the following stages, where:

[0652] - During a first stage, within a time period t i1P the temperature of the nanoparticles increases from a temperature T i to a temperature T iav ,

[0653] - During a second stage, within a time period t i2P the temperature of the nanoparticles remains at the temperature T iav , and

[0654] - During a third stage, within a time period t i3P the temperature of the nanoparticles decreases from T iav to T f .

[0655] The present invention also relates to a purification method according to the present invention, which includes at least one heating step i, where the heating step includes at least one of the following first, second, and / or third stages, where:

[0656] - During a first stage, within a time period t i1P the temperature of the nanoparticles increases from a temperature T i to a temperature T iav ,

[0657] - During a second stage, within a time period t i2P the temperature of the nanoparticles remains at the temperature T iav , and

[0658] - During a third stage, within a time period t i3P the temperature of the nanoparticles decreases from T iav to T f .

[0659] In some cases, T i and / or t i1P is at least 1.0001, 1.1, 1.5, 2, 5, 10, or 100 times lower than T iav and / or t i2P . In some cases, T iav is equal to T1, T2, or T3, and / or t i1P is equal to the time it takes for the temperature to increase to T i . In some other cases, T f and / or t i3P is / are related to T i and / or t i1PThe difference is not greater than 1.0001, 1.1, 1.5, 2, 5, 10, 10 2 or 10 5 times.

[0660] The present invention also relates to a method according to the present invention, preferably a purification method, in which carbon or carbon-containing materials in an amount higher than 10% by mass are removed from the nanoparticles, where the percentage is based on the ratio (%C AT -%C BT ) / %C BT , where %C AT and %C BT are the percentages of carbon or carbonaceous materials before and after treating the nanoparticles with the method, respectively.

[0661] In some cases, (%C AT -%C BT ) / %C BT is greater than 10 -50 , 10 -20 , 10 -5 , 10 -2 , 10 -1 , 1, 5, 10, 50, 75, 90, 95 or 99%. This can be the case when the purification method is effective or when the amount of carbon or carbonaceous materials contained in the nanoparticles before treating the nanoparticles with the purification method is below a certain threshold, preferably below 9, 90, 70, 60, 50, 40, 30, 20, 10 or 1%.

[0662] In some other cases, (%C AT -%C BT ) / %C BT is below 99%, 90%, 70%, 60%, 50%, 40%, 30%, 20%, 10% or 1%. This can be the case when the purification method is not very efficient or when the amount of carbon or carbonaceous materials contained in the nanoparticles before treating the nanoparticles with the purification method is greater than a certain threshold, preferably greater than 99, 90, 70, 60, 50, 40, 30, 20, 10 or 1%.

[0663] In some other cases, (%C AT -%C BT ) / %C BT is between 0.1 and 100, 1 and 99, 10 and 99, 50 and 99, or between 80 and 99%.

[0664] The present invention relates to high-purity iron oxide-based nanoparticles obtained by a method according to the present invention.

[0665] The present invention also relates to high-purity nanoparticles or high-purity nanoparticles that cannot be obtained by this method.

[0666] In one embodiment of the present invention, the high-purity nanoparticles, preferably the coatings of these nanoparticles, comprise: i) 0.8 to 0.999999999 g of iron oxide per gram of nanoparticles, and / or ii) 10 -40 to 10 5 μg of impurities per gram of nanoparticles.

[0667] In one embodiment of the present invention, the high-purity nanoparticles contain less than 90%, 10%, 5%, 2%, preferably 1%, 0.5%, 0.4% or 0.3% by mass of carbon or carbonaceous materials. In some cases, such a low carbon mass percentage enables the nanoparticles to be coated with a coating that does not originate from the cells that generate the nanoparticles.

[0668] In one embodiment of the present invention, the SAR (specific absorption rate) of the high-purity iron oxide nanoparticles is greater than 10 -100 、10 -50 、10 -20 、10 -10 、10 -5 、10 -3 、10 -1 、1、10、10 3 or 10 5 Watts per gram of nanoparticles. In some cases, the SAR of the nanoparticles is the greatest when the amount of impurities in the nanoparticles is the lowest. In some cases, the SAR of the high-purity iron oxide nanoparticles is between 10 -100 and 10 100 、10 -1 and 10 5 、or between 0.1 and 10 3 Watts per gram of nanoparticles. In some cases, the SAR of the nanoparticles is proportional to the slope (preferably the initial slope) of the change in the temperature of the nanoparticles over time (ΔT / δt), preferably when surrounded by, for example, water, biological material, a body part or tissue, where (ΔT / δt) is preferably estimated in °C / sec, where SAR = α(ΔT / δt). In some cases, α = C v / C nano , where C v is the specific heat capacity, preferably of the water, biological material, body part or tissue containing the nanoparticles, and C nano is the nanoparticle concentration or the number or total amount of nanoparticles, preferably which is contained in the water, biological material, body part or tissue. In some cases, the SAR is measured by exposing the high-purity iron oxide nanoparticles to radiation, preferably heat-generating radiation, preferably laser, magnetic field, alternating magnetic field, sound wave, ultrasound, radio frequency.

[0669] In one embodiment of the present invention, the size distribution of high purity iron oxide nanoparticles is less than 10 50 , 10 20 , 10 10 , 10 5 , 10 3 , 10 2 , 10, 1, 10 -1 , 10 -2 or 10 -5 In some cases, when the methods according to the present invention are capable of producing nanoparticles having a low size distribution, the nanoparticle size distribution is low.

[0670] In another embodiment of the present invention, high purity iron oxide nanoparticles, preferably at a concentration greater than 0 -6 , 10 -3 , 10 -1 , 1 or 10 mg nanoparticles per milliliter or per millicubic meter or per cell, destroying more than 1, 10, 10 3 , 10 6 or 10 9 cells.

[0671] The present invention also relates to high purity iron oxide nanoparticles with a concentration of more than 10 -50 , 10 -30 , 10 -10 , 10 -5 , 10 -2 , 10 -1 , 1, 5, 10, 50, 10 2 , 10 3 or 10 5 Milligrams of nanoparticles or milligrams of iron contained in the nanoparticles are preferably generated in a yield per cell, preferably per liter of pre-growth and / or growth medium / media.

[0672] The present invention also relates to nanoparticles obtained by the process according to the invention, wherein the yield of the nanoparticles produced is less than 10 50 , 10 30 , 10 10 , 10 5 , 10 2 , 10, 5, 1, 10 -1 , 10 -2 , 10 -3 or 10 -5 Milligram of nanoparticles or milligram of iron contained in the nanoparticles, preferably per cell, preferably per liter of pre-growth and / or growth medium.

[0673] The present invention also relates to nanoparticles based on high-purity iron oxide according to the invention, wherein the high-purity iron oxide nanoparticles are magnetosomes.

[0674] In one embodiment of the invention, the magnetosomes are nanoparticles produced by magnetotactic bacteria, which are preferably processed after at least one of the following steps: i) the nanoparticles are extracted and / or separated from the bacteria, preferably obtaining magnetosomes comprising crystallized minerals covered by a biofilm, ii) the biofilm is preferably removed using a purification step, iii) the magnetosomes are coated with a coating that does not originate from the nanoparticle producing cells in order to stabilize them, preferably to avoid aggregation and / or deposition of the magnetosomes. The invention also relates to a composition, a medicament, a medical device, a diagnostic composition, a therapeutic composition or a cosmetic composition comprising the high purity iron oxide nanoparticles according to the invention.

[0675] In another embodiment of the invention, the high purity iron oxide nanoparticles produce: i) medical or therapeutic activity, such as by destroying pathological cells, viruses, bacteria, cancer cells, or by being less toxic to healthy cells than pathological cells, viruses, bacteria, cancer cells, ii) diagnostic activity, such as by detecting pathological cells, viruses, bacteria, cancer cells, or by being less toxic to healthy tissue, and / or iii) cosmetic activity, such as by improving a person's appearance.

[0676] In another embodiment of the invention, the high purity iron oxide nanoparticles are non-immunogenic or non-pyrogenic. In this case, they preferably: i) attract or cause the appearance of a small number of immune cells, preferably less than 1, 5, 10, 10 3 , 10 10 , 10 50 or 10 100 immune cells and / or ii) produce an increase in the temperature of the organism below 10 5 , 10 3 , 10 2 , 50, 20, 10, 5, 2, 1 or 0.1C.

[0677] The invention further relates to nanoparticles according to the invention, preferably nanoparticles based on high purity iron oxide, for use in the treatment of diseases, which are preferably selected from: i) diseases associated with cell proliferation which differs from cell proliferation in a healthy individual, ii) diseases associated with the presence of pathological cells, such as tumors or cancer cells in a body part or an individual, iii) diseases associated with the presence of a pathological site, i.e. a site in an individual or a body part containing pathological cells iv) a disease or condition or dysfunction of a body site, v) a disease associated with the presence of radiation-resistant or sound-resistant or laser-resistant or magnetic-resistant cells, vi) infectious diseases, vii) autoimmune diseases, viii) neuropathologies, ix) cancer, x) tumors, xi) diseases or tumor cells containing or caused by at least one cancer, xii) skin diseases, xiii) endocrine diseases, xiv) eye diseases or disorders, xv) intestinal diseases, xvi) communication disorders, xvii) genetic diseases, xviii) nervous system diseases, xix) speech disorders, xx) vulvovaginal diseases, xxi) liver diseases, xxii) heart diseases, xxiii) heating disorders, xxiv) mood disorders, xxv) anemia, preferably iron deficiency, xxvi) personality disorders, xxvii) AIDS, especially neurological aids, xxviii) Parkinson's disease, xxix) Alzheimer's disease, xxx) bacterial and / or fungal infections or contaminations, xxxi) blood diseases, for example due to the absence or lack of effective coagulation, and xxxii) diseases due to immune deficiencies or immune diseases.

[0678] In one embodiment of the present invention, the cancer or tumor is selected from the group consisting of organ cancer, blood cancer, biological system cancer, adrenal cancer, anal cancer, bile duct cancer, bladder cancer, bone cancer, brain cancer, breast cancer, cervical cancer, colon / rectum cancer, endometrial cancer, esophageal cancer, eye cancer, gallbladder cancer, heart cancer, kidney cancer, laryngeal and hypopharyngeal cancer, leukemia, liver cancer, lung cancer, nasal and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin's lymphoma, oral and oropharyngeal cancer, osteosarcoma cancer, ovarian cancer, pancreatic cancer, pancreatic Cancers such as penile cancer, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, skin cancer, small intestine cancer, stomach cancer, testicular cancer, thymic cancer, thyroid cancer, uterine cancer, uterine sarcoma cancer, vaginal cancer, vulvar cancer, Waldenstrom disease, Castle disease, Ewing family of tumors, gastrointestinal carcinoid, gastrointestinal stromal tumor, myelodysplastic syndrome, pituitary tumor, and gestational trophoblastic disease, Hodgkin's disease, Kaposi's sarcoma, malignant mesothelioma and multiple myeloma.

[0679] In another embodiment of the present invention, the use of the nanoparticles according to the present invention for disease treatment occurs or is activated when the nanoparticles are exposed to radiation, and preferably is not initiated or activated when the nanoparticles are not exposed to radiation.

[0680] In another embodiment, the radiation is a laser, acoustic waves such as ultrasound, X-rays, gamma rays, and / or a magnetic field, preferably an alternating magnetic field.

[0681] In another embodiment, the power or intensity of the radiation is between 10 -50 、10 -20 、10 -10 、10 -5 、10 -1 、0, 1 mT, T, W, W / cm, W / cm 2 or W / cm 3 and 1, 5, 10, 10 3 、10 5 、10 10 、10 20 or 10 50 mT, T, W, W / cm, W / cm 2 or W / cm 3 .

[0682] In another embodiment of the present invention, the properties or characteristics of the nanoparticles or methods, preferably described in each individual embodiment or part or sentence of this patent application, can be combined to produce properties or characteristics, preferably nanoparticles or methods.

[0683] In yet another embodiment of the present invention, when a compound such as a nanoparticle or a chemical element has a property that is α times higher, longer, or greater in condition 1 (P1) than in condition 2 (P2), this means that P1 = αP2 or P1 = α + P2, where α is preferably a number or integer greater than 1 or 0.

[0684] In yet another embodiment of the present invention, when a compound such as a nanoparticle or a chemical element has a property that is α times smaller or shorter in condition 1 (P1) than in condition 2 (P2), this means that P1 = P2 / α or P1 = P2 - α, where α is preferably a number or integer greater than 1 or 0.

[0685] The present invention is further disclosed by the following non-limiting embodiments.

[0686] Example

[0687] Materials and Methods

[0688] Measurement of the optical density of all magnetotactic bacteria suspensions used to evaluate bacterial growth: The optical density of different suspensions of magnetotactic bacteria was measured at 565 nm and named OD 565nm , using a Secomam UviLine9400 spectrophotometer. The value of OD 565nm is proportional to the concentration of bacteria in the suspension.

[0689] Observing living magnetotactic bacteria under an applied magnetic field using an optical microscope and measuring their magnetic response: Centrifuge 1 mL of the MSR-1 magnetotactic bacteria suspension at 14,500 rpm for 10 minutes. Remove the growth medium and replace it with a certain volume of PBS 0.1X to achieve an OD 565nm of 0.5. Place 1 μL of this MSR-1 magnetotactic bacteria suspension on a parallel microscope slide (Menzel- 24 mm × 60 mm, 0.13 - 16 mm thickness), and perform microscopic observation using a Zeiss Primo Vert optical microscope with a magnification of 40x. Place four small cube neodymium magnets (Supermagnet, N42W-10-N 10x10x10 mm) with a strength of 1.3 T on the microscope stage and ~2 cm away from the bacteria suspension, such that the magnetic field generated is parallel to the observer's position or parallel to the line between the two binoculars (position 1) or perpendicular to this position (position 2). After positioning the magnets at position 1 or 2 for 20 seconds, estimate the percentage of bacteria aligned in the magnetic field direction by considering 200 magnetotactic bacteria. Bacteria not aligned in the same direction as the magnetic field generated by the magnet are considered non-magnetic. Their number is designated as n BNM . Bacteria aligned in the magnetic field direction generated by the magnet are considered magnetic. Their number is designated as n BM . Then, the percentage of magnetic bacteria is given by n BM / (n BM +n BNM ). The positive magnetic response of magnetotactic bacteria corresponds to n BM / (n BM +n BNM ) > 0.5. The negative magnetic response of magnetotactic bacteria corresponds to n BM / (n BM +n BNM ) < 0.5. In some cases, the percentage of magnetic bacteria can be measured by optical observation under a magnetic field using a microscope.

[0690] Measurement of intracellular iron concentration: The iron concentration within magnetotactic bacteria is determined by a destructive iron assay. For this purpose, centrifuge 2 mL of MSR-1 magnetotactic bacteria at 14,500 g for 10 minutes. Then wash the bacterial pellet twice with 1X PBS and MilliQ water. After the second wash, collect the bacterial pellet and add 1 mL of 12 N hydrochloric acid (HCl) to the pellet under a chemical hood. Heat the sample at 75 °C for 2 hours with stirring at 300 rpm to convert intracellular iron into Fe 3+ and Fe 2+ ions. Then, use hydrogen peroxide (H2O2) at 20% to convert Fe 2+The ion is oxidized to Fe 3+ The presence of Fe ions is shown by adding potassium thiocyanate (KCN, 2 mol / L) in an acidic medium, which results in the formation of a red-orange solution, the color of which depends on the concentration of Fe in the sample 3+ ions. Once KCN is added, the absorbance of the solution is measured at 476 nm. Then the determined relationship between the absorbance value measured at 476 nm and the concentration in iron(III) chloride is used to estimate the iron concentration in the sample. This method can estimate the total intracellular iron concentration 3+

[0691] The elemental chemical composition of magnetosomes is analyzed by ICP-AES: After fermentation, MSR-1 magnetotactic bacteria are concentrated in a 5 L volume by tangential flow filtration to achieve an optical density of 25 - 30. Then the bacteria are dissolved in 1 M KOH solution within 1 hour with stirring at 150 rpm and a temperature of 80 °C. The bacterial lysate containing magnetosomes is placed on a neodymium magnet for 12 hours. Then the magnetosomes are separated from the bacterial lysate and resuspended in 10X PBS. This washing procedure is repeated twice with 10X PBS and three times in MilliQ water. Then the magnetosomes are freeze-dried and heated in a muffle furnace under the following conditions to obtain magnetosome powder containing high-purity iron oxide crystals with a low carbon content. For the analysis of the elemental chemical composition, a solution of 500 μg of this powder is mixed with 200 μ of 12N HCl and 10 ml of 2% filtered HNO3. ICP-AES measurement of the powder gives the amounts of chemical elements contained in the magnetosomes, in μg of these chemical elements per gram of iron contained in the magnetosomes (Ag, Al, As, Ba, Cd, Co, Cr, Cu, Mn, Mo, Ni, Pb, Sb, Se, Si, Sn, Ti, Tl, W, and Zn).

[0692] Chemical products for preparing the growth medium: Potassium alum (AlK(SO4)2·12H2O, reference NFGA6435, Merck); Ammonium hydroxide (NH4OH, reference NFG 1336 - 21 - 6, Acros Organics; reference FG 105422, Merck); Ammonium chloride (NH4Cl, ref.FNG A9434, Merck; ref.FG 1011420001, Merck); Ammonium sulfate ((NH4)2SO4, reference NFG A4418); Biotin ((C 10 H 16 ​N2O3S, referring to NFG B4639, Merck; boric acid (H3BO3, referring to NFG B6768, Merck); calcium chloride (CaCl2, referring to FNG 223506, Merck; referring to FG1.42002, Merck); calcium pantothenate (HOCH2C(CH3)2CH(OH)CONHCH2CH2CO2·1 / 2Ca, referring to FG C0400000, Merck); cobalt(II) nitrate hexahydrate (cobalt(II) hexahydrate hexahydrate, referring to FG 239267, Merck); copper(II) sulfate pentahydrate (CuO4S.5H2O) (referring to NFG C8027, Merck), DL-methionine (CH3SCH2CH2CH(NH2)COOH, referring to NFG M2768, Merck), DL-tryptophan (C11H12N2O2, referring to NFG T3300, Merck); EDTA ((HO2CCH2)2NCH2CH2N(CH2CO2H)2, referring to NFG E6758, Merck); ferric citrate (C6H5FeO7, referring to NFG F3388, Merck; referring to FG B301, Merck); folic acid (C 19 H 19 N7O6, referring to NFG F7876, Merck; referring to FG F0300000, Merck); inositol (C6H 12O6, reference FG PHR1351, Merck); iron(II) sulfate heptahydrate (FeO4S.7H2O, reference NFG F8633, Merck; reference FG 1.03963, Merck); iron(III) oxalate hexahydrate (Fe2(C2O4)3.6H2O, reference NFG381446, Merck); L-histidine (C6H9N3O2, reference FG PHR1108, Merck); magnesium sulfate heptahydrate (MgSO4.7H2O, reference NFG 63138, Merck; reference FG 105882, Merck); manganese(II) sulfate monohydrate (MnO4S.H2O, reference NFG M7899, Merck); nickel(II) chloride hexahydrate (Cl2Ni.6H2O, reference NFG N6136, Merck); nicotinic acid (C6H5NO2, reference NFG N4126, Merck); nitrilotriacetic acid trisodium salt ((C6H6NO6Na3, reference NFGN0253, Merck); p-aminobenzoic acid (H2NC6H4CO2H, reference NFG A9878, Merck); dipotassium hydrogen phosphate (K2HPO4, reference NFG P8281, Merck; reference FG 105101, Merck); potassium dihydrogen phosphate (KH2PO4, reference NFGP9791, Merck); protoporphyrin IX (C 34 H 34 N4O4, ref. NFG P8293, Merck); pyridoxine hydrochloride (C 12 H 17 ClN4OS.HCl, ref. FNG P9755, Merck); riboflavin (C 17 H 20 N4O6, reference NFG R9504, Merck; reference FG PHR1054, Merck); sodium chloride (NaCl, ref. FNG S7653, Merck); sodium lactate (C3H5NaO3, reference NFGL1375, Merck; reference FG 106522, Merck); sodium molybdate dihydrate (Na2Mo4.2H2O, reference NFG M1003, Merck); sodium selenite pentahydrate (Na2SeO3.5H2O, reference FG 89771, Merck); thiamine HCL (C 12 H 17ClN4OS.HCl, ref. FNG47858, Merck, ref. FG PHR1037, Merck); yeast extract (see NFGY1625, Merck); zinc sulfate heptahydrate (O4SZn.7H2O, ref. NFG Z0251, Merck). NFG refers to non-pharmaceutical grade chemicals used for preparing growth media; FG designates pharmaceutical grade chemicals used for preparing growth media. We also use deionized water (H2O) with a resistivity of 15 MΩ.

[0693] Composition of different mineral elixirs: The composition of different mineral elixirs (V0, CB1, V2, CB2, CB3, CB4, CB5, CB7, CB9, CB10, CB11, CB12, CB13) is shown in Table 6, where the quantities (grams) represent the different chemicals used to prepare 1 liter of these elixirs.

[0694] Composition of different yeast extracts: The composition of different yeast extracts (YE, YNBWAA, YNBWoAA, YNBWoAA.AS) is given in Table 7, where the amounts (in grams) of the different chemicals used to prepare 1 liter of these yeast extracts are given. YNBWAA, YNBWoAA, YNBWoAA.AS represent reduced yeast extracts, while YE represents non-reduced yeast extract (ref: Y0875, Sigma). YE contains nitrogenous compounds, carbon, sulfur, micronutrients, vitamin B complex, and other important growth factors.

[0695] Composition of different vitamin cocktails: The composition of different vitamin cocktails (Vit1X, Vit5X, Vit10X, Vit0.5X, Vit0.1X) is given in Table 8, where the amounts (in grams) of the different chemicals used to prepare 1 liter of these vitamin cocktails are given.

[0696] Composition of the pre-growth medium for Condition 1 (Table 1): 1 liter of the pre-growth medium contains 1 liter of deionized water, 2.6 g of sodium lactate, 0.4 g of ammonium chloride, 0.1 g of magnesium sulfate heptahydrate, 0.5 g of dipotassium hydrogen phosphate, 0.1 g of yeast extract YE (Table 7), and 0.5 ml of any one of the mineral elixirs V0, CB1, V2, CB2, CB3, CB4, CB5, CB7, CB9, CB10, CB11, CB12, or CB13 (Table 6).

[0697] Composition of the growth medium for Condition 1 (Table 1): 1 liter of growth medium contains 1 liter of deionized water, 2.6 g of sodium lactate, 0.4 g of ammonium chloride, 0.1 g of magnesium sulfate heptahydrate, 0.5 g of dipotassium hydrogen phosphate, 0.1 g of yeast extract YE (Table 7), 0.5 mL of any one of the mineral elixirs V0, CB1, V2, CB2, CB3, CB4, CB5, CB7, CB9, CB10, CB11, CB12 or CB13 (Table 6) and 10 mL of iron citrate (initial concentration 20 mM).

[0698] Composition of the pre-growth medium for Condition 2 (Table 2): 1 liter of pre-growth medium contains 1 liter of deionized water: 2.6 g of sodium lactate, 0.4 g of ammonium chloride, 0.1 g of magnesium sulfate heptahydrate, 0.5 g of dipotassium hydrogen phosphate, 0.1 g of yeast extract YE, any one of YNBWAA, YNBWoAA, YNBWoAA.AS (Table 7) and 0.5 mL of the mineral elixir CB3 (Table 6).

[0699] Composition of the growth medium for Condition 2 (Table 2): 1 liter of growth medium contains 1 liter of deionized water, 2.6 g of sodium lactate, 0.4 g of ammonium chloride, 0.1 g of magnesium sulfate heptahydrate, 0.5 g of dipotassium hydrogen phosphate, 0.1 g of yeast extract YE, any one of YNBWAA, YNBWoAA or YNBWoAA.AS (Table 7), 0.5 mL of the mineral elixir CB3 (Table 6) and 10 mL of iron citrate (initial concentration 20 mM).

[0700] Composition of the pre-growth medium for Condition 3 (Table 3): 1 liter of pre-growth medium contains 1 liter of deionized water, 2.6 g of sodium lactate, 0.4 g of ammonium chloride, 0.1 g of magnesium sulfate heptahydrate, 0.5 g of dipotassium hydrogen phosphate, 0.1 m of any one of vitamin Vit1X, Vit5X, Vit10X, Vit5X, Vit0.5X or Vit0.1X (Table 8) and 0.5 mL of the mineral elixir CB3 (Table 6).

[0701] Composition of the growth medium for Condition 3 (Table 3): 1 liter of growth medium contains 1 liter of deionized water, 2.6 g of sodium lactate, 0.4 g of ammonium chloride, 0.1 g of magnesium sulfate heptahydrate, 0.5 g of dipotassium hydrogen phosphate, 0.1 mL of any one of the vitamin cocktails Vit1X, Vit5X, Vit10X, Vit5X, Vit0.5X or Vit0.1X (Table 8), 0.5 mL of the mineral elixir CB3 (Table 6) and 10 mL of iron citrate (initial concentration 20 mM).

[0702] Composition of the pre-growth medium for Condition 4 (Table 4): 1 liter of pre-growth medium contains 1 liter of deionized water, 2.6 g of sodium lactate, 0.4 g of ammonium chloride, 0.1 g of magnesium sulfate heptahydrate, 0.5 g of dipotassium hydrogen phosphate, 0.1 mL of any one of the individual vitamins Bt, CP, FA, I, NA, AA, P, R, or T (Table 9), and 0.5 mL of mineral elixir CB3 (Table 6).

[0703] Composition of the growth medium for Condition 4 (Table 4): 1 liter of growth medium contains 1 liter of deionized water, 2.6 g of sodium lactate, 0.4 g of ammonium chloride, 0.1 g of magnesium sulfate heptahydrate, 0.5 g of dipotassium hydrogen phosphate, 0.1 mL of any one of the individual vitamins Bt, CP, FA, I, NA, AA, P, R, or T (Table 9), 0.5 mL of mineral elixir CB3 (Table 6), and 10 mL of iron citrate (20 mM initial concentration).

[0704] Composition of the pre-growth medium for Condition 5 (Table 5): Table 5 gives the compositions of different pre-growth media with different concentrations of the main components of the pre-growth medium, namely sodium lactate, ammonium chloride, magnesium sulfate heptahydrate, dipotassium phosphate (N, SL0, SL0.5X, SL0.2X, SL0.1X, AC0, AC0.5X, AC0.2X, AC0.1X, MG0, MG0.5X, MG0, MG0.5X, MG0.2X, MG0.1X, P, P0.5X, P0.2X, P0.1X), where the amounts (in grams) of the different chemicals used to prepare 1 liter of these pre-growth media are shown.

[0705] Compositions of the pre-growth medium, growth medium, and fed-batch medium for Condition 6 prepared using non-pharmaceutical grade chemicals (Table 14(a)): The pre-growth media B1 and B4 contain 2.6 g of sodium lactate, 0.4 g of ammonium chloride, 0.1 g of magnesium sulfate heptahydrate, 0.5 g of dipotassium hydrogen phosphate, 0.1 mL of vitamin mixture Vit 0.1X (Table 8), and 0.5 mL of mineral elixir CB3 (Table 6) in 1 liter of deionized water. The growth media B1 and B4 contain 104 g of sodium lactate, 16 g of ammonium chloride, 1.2 g of magnesium sulfate heptahydrate, 2.8 g of dipotassium hydrogen phosphate, 3.2 mL of vitamin mixture Vit0.1X (Table 8)), and 2.8 mL of mineral elixir CB3 (Table 6) in 1 liter of deionized water. The fed-batch media B1 and B4 contain 100 g of lactic acid, 4.8 g of ammonia, 6 g of dipotassium hydrogen phosphate, 2.4 g of magnesium sulfate heptahydrate, 1 mL of vitamin mixture Vit0.1X (Table 8), 7 mL of mineral elixir CB3 (Table 6), and 1.8 g of iron citrate (B1) or 2 g of iron(III) chloride (B4) in 1 liter of water.

[0706] Pre-growth medium for Condition 6, composition of growth medium and fed-batch medium prepared using pharmaceutical-grade chemicals (Table 14(b)): Pre-growth media B2 and B3 contain, in 1 liter of deionized water, 2.6 g of sodium lactate, 0.4 g of ammonium chloride, 0.1 g of magnesium sulfate heptahydrate, 0.5 g of dipotassium hydrogen phosphate, 0.1 mL of vitamin mixture Vit 0.1X (Table 8), and 0.5 mL of mineral elixir CB3 (Table 6). Growth media B1 and B4 contain, in 1 liter of deionized water, 104 g of sodium lactate, 16 g of ammonium chloride, 1.2 g of magnesium sulfate heptahydrate, 2.8 g of dipotassium hydrogen phosphate, 3.2 mL of vitamin mixture Vit 0.1X (Table 8), and 2.8 mL of mineral elixir CB3 (Table 6). Fed-batch media B1 and B4 contain, in 1 liter of water, 100 g of lactic acid, 4.8 g of ammonia, 6 g of dipotassium hydrogen phosphate, 2.4 g of magnesium sulfate heptahydrate, 1 mL of vitamin mixture Vit 0.1X (Table 8), 7 mL of mineral elixir CB3 (Table 6), and 1.8 g of iron citrate (B2) or 2 g of iron chloride (B3).

[0707] Stock of Magnetospirillum magneticum strain MSR-1 for different culture media: Magnetospirillum magneticum strain MSR-1 is commercialized by DSMZ under the designation DSM 6361. After receipt, the MSR-1 bacterial suspension is stored at an OD of 0.01 565nm (optical density measured at 565 nm), corresponding to a bacterial concentration of 5.10 7 strains per milliliter of medium (medium DSMZ380 for culturing Magnetospirillum magneticum strain DSMZ 6361), added (5 mL of bacterial suspension per tube) or to 1.5 mL Eppendorf tubes (600 μL of bacterial suspension per tube). The suspension of MSR-1 bacteria is stored in a -80 °C freezer to constitute the cell stock. In some cases, the culture medium and / or pre-growth medium can be the same as the growth and / or pre-growth medium.

[0708] Note: The number X after D in DX represents the number of days after the start of the pre-growth step, preferably the day when the magnetotactic bacteria are first added to the pre-growth medium or the first sub-step of the pre-growth step.

[0709] Example 1: Determination of the minimum mineral elixir capable of bacterial growth and magnetosome synthesis: This example describes an experimental protocol for minimizing the mineral elixir composition while enabling the growth of MSR-1 magnetotactic bacteria and the synthesis of magnetosomes by these cells. In this example, non-pharmaceutical grade chemicals were used to prepare the growth medium. The composition (Condition 1) of 1 liter of pre-growth and growth medium used in this example is shown in Table 1. On the first day of the experiment (D1), the first step involved removing a 15 mL test tube containing a 5 mL MSR-1 cell stock tube from the refrigerator at -80 °C and allowing it to thaw at room temperature for 10 minutes. In a fume hood, we collected 100 μl containing 5.10 6 MSR-1 magnetotactic bacteria and placed them in a 50 mL tube containing 8 mL of pre-growth medium that had been filtered. Overall, 13 different culture conditions were tested corresponding to 13 different mineral elixirs. The 50 mL test tubes were placed in an incubator at 29.5 °C and cultured for 6 days between D1 and D6 under shaking conditions of 150 rpm. The second step involved adding an iron source to the growth medium to enable the MSR-1 bacteria to synthesize magnetosomes. After 6 days of pre-growth on D6, the 50 ml test tubes were placed in a fume hood. Then, 30 ml of filtered medium was added to this 50 mL tube, and the bacteria grew between D6 and D13. A positive magnetic response at D13 and a ratio of the optical density at D13 to the optical density at D6 greater than 1 were observed in V0, V2, CB2, CB3, CB4, CB5, CB7, CB10, CB11, CB12, CB13 (Condition 1). In contrast, no magnetic response was observed under Conditions CB1 and CB9, where the concentration of the chemical elements constituting the mineral elixir was less than 10 -5 g / L. In summary, the minimum mineral elixir capable of enabling a significant increase in optical density (OD 565nmD13 / OD 565nmD6 greater than 4.8) and magnetosome synthesis (positive magnetic response) for the growth of MSR-1 bacteria is CB13, which consists only of ferrous sulfate heptahydrate at a concentration of 1 gram per liter and calcium chloride at a concentration of 20 grams per liter.

[0710] Example 2: Determination of a growth medium for the growth of magnetotactic bacteria and magnetosome synthesis without yeast extract: This example describes an experimental protocol for determining a reducing medium to replace yeast extract that enables the growth of Magnetospirillum magneticum strain AMB-1 (MSR-1) magnetotactic bacteria and the synthesis of magnetosomes using these bacteria. In this example, we used non-pharmaceutical grade chemicals to prepare the growth medium. The composition of the pre-growth and growth media in 1 liter of deionized water is shown in Tables 2 (Condition 2), 3 (Condition 3), and 4 (Condition 4). On the first day of the experiment (D1), the first step was to collect a 15 ml tube containing a 5 mL MSR-1 cell stock tube from the refrigerator at -80 °C and thaw the tube by placing it at room temperature for 10 minutes. In a fume hood, we collected 100 μl containing 5.10 6 MSR-1 magnetotactic bacteria and placed them into a 50 mL tube filled with 8 mL of pre-growth medium that had been filtered (Condition 2 (Table 2), Condition 3 (Table 3)), or Condition 4 (Table 4)). The 50 mL test tube was placed in an incubator at 29.5 °C and cultured for 6 days between D1 and D6 under shaking conditions of 150 rpm. The second step involved adding an iron source to the growth medium to enable the MSR-1 bacteria to synthesize magnetosomes. After 6 days of pre-growth on D6, the 50 ml test tube was placed in a fume hood, and 30 ml of filtered growth medium was added to the 50 mL tube (Condition 2, Table 2, Condition 3, Table 3, Condition 4, Table 4), and the bacteria grew between D6 and D13. Tables 11 and 12 show that for yeast extract YE, YNBWAA, YNBWAA, YNBWoAA, YNBWoAA, YNBWoAA.AS (Condition 2), for Vit1X, Vit0.5X, Vit0.1X, (Condition 3), biotin (Bt), folic acid (FA), nicotinic acid (NA), riboflavin (R), thiamine (T)) (Condition 4), the magnetic response at D13 was greater than 90% and the ratio of the optical density measured at D13 to the optical density measured at D6 was greater than 1. In contrast, for Conditions Vit5X, Vit10X (Condition 3), the magnetic response was 0, and for Conditions CP, I, AA, P (Condition 4), the magnetic response was very low. In summary, yeast extract can be replaced with single vitamins: biotin, folic acid, riboflavin, nicotinic acid, or thiamine. The OD 565nmD13 / OD 565nmD6 values for these vitamins were 9.8 (biotin), 2.9 (folic acid), 4.8 (riboflavin), 2.4 (nicotinic acid), 5.8 (thiamine), and a magnetic response of 90% (Table 11).

[0711] Example 3: Determine the minimum concentrations of the main components of the growth medium (sodium lactate, ammonium chloride, magnesium sulfate, potassium phosphate) to enable the growth of magnetotactic bacteria and the synthesis of magnetosomes using these bacteria. This example describes the experimental protocol for determining the reduced growth medium that enables the growth of MSR-1 magnetotactic bacteria and the synthesis of magnetosomes using these bacteria. In this example, we used non-pharmaceutical grade chemicals to prepare the growth medium. We varied the concentration of sodium lactate (conditions SL0, SL0.5X, SL0.2X, SL0.1X), ammonium chloride (AC0, AC0.5X, AC0.2X, AC0.1X), magnesium sulfate heptahydrate (MG0, MG0.5X, MG0.2X, MG0.1X), and dipotassium hydrogen phosphate (P0, P0.5X, P0.2X, P0.1X). Table 5 summarizes the chemical composition and concentration of 1 liter of pre-growth medium and growth media N, SL0, SL0.5X, SL0.2X, SL0.1X, AC0, AC0.5X, AC0.2X, AC1 liter 0.1X, MG0, MG0.5X, MG0.2X, MG0.1X, P0, P0.5X, P0.2X, P0.1X. On the first day of the experiment (D1), the first step was to collect a 15 ml test tube containing a 5 mL MSR-1 cell stock tube from the refrigerator at -80 °C and thaw the test tube by placing it at room temperature for 10 minutes. In the fume hood, we collected 100 μl containing 5.10 6 MSR-1 magnetotactic bacteria from these tubes and placed them in a 50 mL tube filled with 8 mL of pre-growth medium that had been filtered (condition 5 (Table 5)). The 50 mL test tube was placed in an incubator at 29.5 °C and cultured for 6 days between D1 and D6 under shaking conditions of 150 rpm. The second step involved adding an iron source to the growth medium to enable the MSR-1 bacteria to synthesize magnetosomes. After 6 days of pre-growth on D6, the 50 ml test tube was placed in the fume hood, and 30 ml of filtered growth medium was added to the 50 mL tube, and the bacteria grew between D6 and D13. For conditions N, P0.5X, P0.2X, the growth ratio or the ratio between the optical density measured at D13 and the optical density measured at D6 was greater than 1, and a positive magnetic response (magnetic response > 90%) was observed at D13. This indicates that these conditions enabled the bacteria to grow and produce magnetosomes. In contrast, for conditions SL0, SL0.5X, SL0.2X, SL0.1X, AC0, AC0.5X, AC0.2X, AC0.1X, MG0, MG0.5X, MG0.2X, MG0.1X, P0.1X, P0, the synthesis of magnetosomes was very low (magnetic response < 50%). In summary, the concentration of potassium phosphate can be reduced by 2 or 5 times in the growth medium without affecting the growth and production of magnetosomes. In fact, the OD 565nmD13 / OD 565nmD6The values are 1.5 (condition P0.5X), 2.1 (condition P0.2X), and the percentage of positive magnetic response between bacteria is greater than 90% (Table 13). On the contrary, the concentrations of other chemicals in the culture medium (ammonium chloride, sodium lactate, magnesium phosphate) cannot be reduced without significantly affecting the growth and / or magnetic response of MSR-1 magnetotactic bacteria.

[0712] Example 4: Determine the iron source in a 1-liter fermenter for the growth of magnetotactic bacteria and the synthesis of magnetosomes using these bacteria, and reduce the acquisition of impurities by preparing pre-growth and / or growth media using high-purity pharmaceutical-grade chemical products. This example describes the experimental protocol for determining the iron source for the growth of MSR-1 magnetotactic bacteria and the synthesis of magnetosomes using these bacteria, and reducing the impurities contained in the obtained magnetosomes by using pharmaceutical-grade chemicals (condition 6). In this example, we used pharmaceutical-grade chemicals to prepare B2 and B3 growth media (Table 14(b)) and non-pharmaceutical-grade chemicals to prepare B1 and B4 growth media (Table 14(a)). The compositions of 1-liter pre-growth media, growth media, and fed-batch media are shown in Tables 14(a) and 14(b). On the first day (D1), the first step of pre-growth included collecting 1 tube of 1.5 mL Eppendorf containing 600 μL of MSR-1 cell stock from a -80 °C refrigerator and thawing the tube by placing it at room temperature for 10 minutes. In a fume hood, we collected 300 μl containing 1.5×10 7 MSR-1 magnetotactic bacteria from these tubes and placed them in a 500 mL sterile bottle filled with 250 mL of filtered pre-growth medium. The 500 ml bottle was cultured in an incubator at 29.5 °C for 7 days between D1 and D7.

[0713] Then, the second step of pre-growth is carried out in a larger 2L bottle. Seven days after pre-growth at D8, the 500mL bottle is placed in a fume hood. The pre-growth medium containing MSR-1 bacteria is manually transferred to a 2L sterile bottle containing 1.5L of filtered pre-growth medium for the second step of pre-growth. The 2L bottle is cultured in an incubator at 29.5°C under shaking conditions of 150 rpm between D8 and D9 for 1 day. On the 9th day (D9), the growth step begins. For this, four fermenters (conditions, B1, B2, B3, B4) of 1.5L are filled with 780mL of deionized water and autoclaved. Then, 20mL of filtered growth medium is loaded into the fermenters. Then, each of the four fermenters (conditions, B1, B2, B3, B4) is filled with 200mL of pre-growth medium containing MSR-1 bacteria derived from the second step of pre-growth. Between D9 and D11, an acidic fed-batch medium containing an iron source is added to the growth medium to enable MSR-1 bacteria to synthesize magnetosomes while maintaining the pH of the growth medium at 6.9. During the growth step, the temperature is maintained at 29.5°C, the airflow is maintained at 0.05mL / min, and stirring is carried out at 200rpm. Table 15 shows the optical density measured at 565nm of the bacterial suspension on different days during the pre-growth step (D0 and D8) and the growth step (D9, D10, D11) for conditions B1, B2, B3, and B4. After fermentation at D13, the MSR-1 cells from fermenters B1, B2, B3, and B4 (conditions B1 to B4) are concentrated by centrifugation at 4000rpm for 45 minutes. To lyse the bacteria, the MSR-1 cells from fermenters B1, B2, B3, B4 are resuspended in 15mL of 1M KOH solution and heated in a 20mL glass bottle in an ultrasonic cleaning tank at 25kHz and 80°C for 2 hours. After bacterial lysis, the magnetosomes from the MSR-1 cells are separated from the organic materials overnight using a neodymium magnet. On D14, the magnetosomes from conditions B1, B2, B3, B4 are washed twice with 15mL of 10X phosphate-buffered saline and 15mL of deionized water using a 15mL neodymium magnet. During each wash, the magnetosome suspension is placed next to the neodymium magnet for 2 hours to attract the magnetosomes. The supernatant containing organic impurities is discarded and replaced with 15mL of 10X phosphate-buffered saline or 15mL of deionized water. On D16, after the last wash, the supernatant is discarded, and the magnetosomes from conditions B1, B2, B3, B4 are transferred to a ceramic cup and dried overnight by placing them next to a neodymium magnet. On D17, the remaining liquid is discarded, and the magnetosomes are placed in a ceramic cup and heated in a muffle furnace at 200°C for 30 minutes, at 300°C for 1 hour, and at 380°C for 1 hour. On D17, ~1mg of purified magnetosomes from conditions B1, B2, B3, B4 are injected into a 15mL tube filled with 200μL of 12N HCL.A 15 mL test tube containing magnetosomes was vortexed and incubated at room temperature for 2 hours, and then filled with 9.8 mL of 2% HNO3. Thereafter, the μg elemental impurity concentration per gram of nanoparticles was measured by ICP-AES. For conditions B1, B2, B3, B4, the results of these measurements are indicated in Table 16, where the elemental impurities are Ag (silver), Al (aluminum), As (arsenic), Ba (barium), Cd (cadmium), Co. (cobalt), Cr (chromium), Cu (copper), Mn (manganese), Mo (molybdenum), Ni (nickel), Pb (lead), Sb (antimony), Se (selenium), Si (silica), Sn (tin), Ti (titanium), Tl (tall), W (tungstate), Zn (zinc). In summary, condition B3 produced OD. 565nmD11 / OD 565nmD9 The maximum value of 26.8 and a certain percentage of positive magnetic responses in bacteria (>90%) (Table 15) indicate that iron(III) chloride is the best iron source. In addition, except for Pb, the elemental impurity concentration is reduced in condition B3 using pharmaceutical-grade chemicals compared to condition B4 using non-pharmaceutical-grade chemicals (Table 16).

[0714] Example 5 (Purification Method):

[0715] Materials and Methods: Note: In this example, weight can be replaced by mass, preferably resulting in the same meaning.

[0716] Equipment for analyzing and heating various samples:

[0717] TGA-DSC : "Thermogravimetric analysis" (TGA) combined with "differential scanning calorimetry" (DSC) was used to measure the heat flow (mW) or the percentage of mass loss of powders containing freeze-dried magnetosomes (treated or untreated) or freeze-dried whole bacteria or freeze-dried SIGMA nanoparticles as a function of the heating temperature of these powders. The powders were heated at a rate of 6 °C per minute between 20 °C and 600 °C for measurement. The derivative of the mass change of the powders was plotted as a curve as a function of temperature. The TGA-DSC curve can define the temperature at which the materials, preferably organic materials, located in or on the magnetosomes or nanoparticles are degraded, removed from the nanomaterials, or transferred. ATG and DSC analyses were performed using an SDT Q600 (TA Instrument). It consists of a sealed housing, a temperature-controlled furnace, a microbalance, and a thermocouple for measuring temperature. Freeze-dried bacteria, freeze-dried magnetosomes, and SIGMA nanoparticle powders with a mass of 3 mg were used for TGA-DSC analysis.

[0718] CHNS:The "Elemental Carbon, Hydrogen, Nitrogen, and Sulfur Analyzer" CHNS measurements were performed using a CHNS analyzer (Flash Elemental Analyzer EA 1112 from ThermoFischer scientific). Each measurement used 3 mg of freeze-dried magnetosomes (treatment conditions n°1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11), freeze-dried whole bacteria, and freeze-dried SIGMA nanoparticles (untreated). CHNS measurements can determine the mass percentages of carbon and nitrogen in different powders. Powders of freeze-dried bacteria, freeze-dried magnetosomes, and SIGMA nanoparticles with a mass of 3 mg were used for CHNS measurements.

[0719] Furnace: A muffle furnace (Nabertherm L9 / 11 / B410) was used to heat 30 mg or 500 mg of magnetosomes extracted from magnetotactic bacteria under conditions 1 or 2, which were either not heat-treated above 200 °C or heat-treated above 200 °C (following conditions n°3, 4, 5, 6, 7, 8, 9, 10, or 11). For this purpose, the powder of each 30 or 500 mg sample was placed in an uncovered porcelain cup and placed in the center of the furnace. A program was used to perform different heating conditions. The furnace can maintain the temperature of the nanoparticles or the temperature inside the furnace at a given temperature ±2 °C, or the furnace can obtain a stable temperature between 20 °C and 380 °C with a maximum fluctuation of 2 °C.

[0720] Sample containing all magnetotactic bacteria (Sample 0): Magnetotactic bacteria obtained from condition 1 (mineral elixir V2, Table 1) were collected and concentrated using a tangential filtration system to an optical density measured at 565 nm (OD565nm) between 100 and 200. Sample 0 contains all concentrated magnetotactic bacteria.

[0721] The sample contains magnetosomes extracted from magnetotactic bacteria that have never been heat-treated above 200 °C (Sample 1 and 2)：

[0722] Dissolution condition 1 (Sample 1) : 100 ml of a concentration at OD 565nmSample 0 with a size of 120 was mixed with 400 ml of 5M NaOH and heated in an ultrasonic cleaning bath at 60 °C for 1 hour under ultrasonic treatment to dissolve the bacteria. Then, the treated magnetosomes were separated from the bacterial debris by placing a neodymium magnet against the wall of the container containing the dissolved bacterial suspension and replacing the supernatant containing the culture medium and bacterial debris with 1X PBS. The resulting suspension was then ultrasonically treated in 1X PBS at 10 W for 20 seconds, placed against the neodymium magnet for 15 minutes, the supernatant was removed, and the treated magnetosomes were resuspended in 1X PBS. This ultrasonic and magnetic separation was repeated four times. For the entire fermenter, this treatment was repeated 10 times in 10 different volumes. Thereby, a pyrogenic magnetosome chain extracted from the MSR-1 magnetotactic bacterium was obtained, that is, the magnetosomes contained in 1.7 ml of water had approximately 500 mg of iron. Sample 1 contained magnetosomes obtained from dissolution condition 1.

[0723] Dissolution condition 2 (Sample 2) : The concentrated magnetotactic bacteria were frozen at -80 °C for 48 hours. After thawing and diluting the concentrate with MilliQ water to obtain an OD565nm of 30, a certain amount of KOH was added to the concentrated bacteria to obtain a final KOH concentration of 1M. The solution was transferred to a polypropylene (PP) bottle and placed in a water bath at 80 °C while stirring with a mechanical stirring pad (Fisher Scientific) at 150 rpm for 30 minutes. Then, the contents of the bottle were transferred to another 4 2-L glass bottles. Each bottle was placed on a NdFeB magnet for 12 hours to magnetically separate the extracted magnetosomes from the bacterial debris. Then, the magnetosomes were washed 6 times in a 500-mL bottle by magnetic selection until a clear supernatant was obtained. The first two washes were performed with 10X PBS, which could restore the solution to a neutral pH. Then, water was used for the other four washes. After dissolution, the alkaline pH of the lysate caused by KOH was restored to a neutral pH to avoid damaging the magnetosomes. Thereby, a pyrogenic magnetosome chain extracted from strain MSR-1 was obtained, that is, the magnetosomes in 1.7 ml had approximately 500 mg of iron. Sample 2 contained magnetosomes obtained from dissolution condition 2.

[0724] Sample (condition 3) containing magnetosomes extracted from magnetotactic bacteria and treated with phenol-chloroform

[0725] Treatment condition 3 (Sample 3): 100 μl of the iron suspension containing 30 mg of ferromagnetic material obtained under lysis condition 1 was mixed with 200 ml of a solution containing 1% Triton X-100 and 1% SDS. The mixture was heated overnight at 50 °C, placed against a neodymium magnet, the supernatant was removed and replaced with 80 ml of phenol at pH 8. The resulting suspension was heated under sonication at 60 °C for 2 hours and then kept at 60 °C overnight without sonication, placed against a neodymium magnet, the supernatant of the suspension was removed and replaced with 80 ml of chloroform. The suspension containing chloroform was placed against a neodymium magnet, the supernatant was removed, and the residual chloroform adsorbed on the surface of the treated magnetosomes was removed by heating these magnetosomes in a fume hood for 2 hours. Finally, the cores of the magnetosomes were desorbed from the glass walls of the tube containing them by adding 80 ml of 1 M NaOH and heating in an ultrasonic bath at 60 °C for 1 hour. The suspension containing the magnetosome cores was placed against a neodymium magnet. The supernatant was removed and replaced with sterile MilliQ water. The suspension was sonicated at 10 W for 20 seconds. This washing procedure was repeated four times. Purified pyrogen-free magnetosomes were obtained in a small amount of pyrogen-free water. Sample 3 contains magnetosomes obtained after treatment with condition 3.

[0726] The sample contains magnetosomes extracted from magnetotactic bacteria and is heated at a temperature above 200 °C (Samples 4 to 11):

[0727] Heat treatment condition 4 (Sample 4): 100 μl of a magnetosome suspension containing approximately 30 mg of iron extracted from MSR-1 magnetotactic bacteria under dissolution condition 2 was lyophilized, introduced into a porcelain crucible, and baked in a Nabertherm L9 / 11 / B410 furnace. The heating protocol was as follows. The temperature of the furnace was increased from 20 °C to 200 °C at a rate of 6 °C / min until the temperature of the furnace reached 200 °C and the temperature in the furnace was maintained at 200 °C for one hour. Then the furnace temperature was decreased from 200 °C to 25 °C within 12 hours. Sample 4 contains magnetosomes obtained after treatment with condition 4.

[0728] Heat treatment condition 5 (Sample 5): 100 μl of a magnetosome suspension containing approximately 30 mg of iron extracted from MSR-1 magnetotactic bacteria under dissolution condition 2 was lyophilized, introduced into a porcelain crucible, and baked in a Nabertherm L9 / 11 / B410 furnace. The heating protocol was as follows. The temperature of the furnace was increased from 20 °C to 400 °C at a rate of 6 °C / min until the temperature of the furnace reached 400 °C. The temperature inside the furnace was maintained at 400 °C for 1 hour. Then the furnace temperature was decreased from 400 °C to 25 °C wi...

Claims

1. A method for producing highly pure ferromagnetic or ferrimagnetic iron oxide nanoparticles using ferromagnetic or ferrimagnetic iron oxide nanoparticle-producing cells, wherein the nanoparticle-producing cells are magnetotactic bacteria selected from the strains Magnetospirillum magneticum AMB-1, Magnetococcus strain MC-1, three facultative anaerobic vibrio strains MV-1, MV-2, and MV-4, Magnetospirillum magnetotacticum strain MS-1, Magnetospirillum gryphiswaldense strain MSR-1, Magnetospirillum magneticum strain MGT-1, and the obligate anaerobic bacterium Desulfovibrio magneticus RS-1, the method comprising: a) A pre-growth stage, including amplifying the cells producing the ferrimagnetic or ferromagnetic iron oxide nanoparticles in a pre-growth and / or fed-batch medium, and b) A growth stage, including amplifying the cells producing the ferrimagnetic or ferromagnetic iron oxide nanoparticles derived from the pre-growth stage in a growth and / or fed-batch medium, wherein each kilogram or liter of the pre-growth and / or growth and / or fed-batch medium contains: i) Not more than 0.005 grams of yeast extract, and ii) Zero boric acid and nitrilotriacetic acid, which are carcinogenic, mutagenic and reprotoxic (CMR) reagents, wherein, in the pre-growth stage and the growth stage, when the fed-batch medium is present, the fed-batch medium is a medium that supplements the pre-growth and / or growth medium, and wherein, more ferrimagnetic or ferromagnetic iron oxide nanoparticles are produced in the growth stage than in the pre-growth stage, and the method further includes a purification method by removing at least one impurity from the ferrimagnetic or ferromagnetic iron oxide nanoparticles, which includes step 1 and step 2, wherein: - During step 1, the temperature of the ferrimagnetic or ferromagnetic iron oxide nanoparticles is raised to temperature T1 and then maintained at T1 for a heating time of 1 minute to 6 hours, where T1 is between 50°C and 250°C; - During step 2, the temperature of the ferrimagnetic or ferromagnetic iron oxide nanoparticles is raised to temperature T2 and then maintained at T2 for a heating time of 1 minute to 12 hours, where T2 is between 350°C and 500°C.

2. The method according to claim 1, wherein both step 1 and step 2 each comprise at least two stages: (i) The second stage of each of step 1 and step 2, during which the temperature of at least one nanoparticle is maintained at a heating temperature as follows: a) For step 1, between 50 °C and 250 °C; and b) For step 2, between 350 °C and 500 °C; and (ii) At least one other stage selected from: A first stage, during which the temperature of at least one nanoparticle is increased from an initial temperature to the heating temperature of the second stage, wherein the initial temperature is less than 102 °C or less than the heating temperature of the second stage, and A third stage, during which the temperature of at least one nanoparticle is decreased from the heating temperature of the second stage to a final temperature.

3. The method according to claim 1, wherein at least one of the nanoparticles has a crystalline structure, and the heating temperature in the second stage and the final temperature reached by the at least one nanoparticle in the third stage do not induce melting of the at least one nanoparticle, destruction of the at least one nanoparticle, denaturation of the at least one nanoparticle, or modification of the crystalline structure of the at least one nanoparticle, the modification of the crystalline structure of the at least one nanoparticle being different from the modification caused by oxidation of the at least one nanoparticle or different from a change in the oxidation state of the at least one nanoparticle, wherein the at least one nanoparticle has at least one property selected from the group consisting of magnetic, diamagnetic, ferromagnetic, ferrimagnetic, and paramagnetic, and at least one impurity is removed from the at least one nanoparticle by the method.

4. The method according to claim 1, wherein the culture medium is around the magnetosome.

5. The method according to claim 1, wherein the growth stage is distinguished from the pre-growth stage by at least one characteristic selected from the group consisting of: i) The ratio of C FeGS / C FePGS is greater than 1, where C FeGS and C FePGS are the concentrations of iron or an iron source in the growth medium and the pre-growth medium, respectively, ii) The ratio of C CGS / C CPGS is greater than 1, where C CGS and C CPGS are the concentrations of carbon or a carbon source in the growth medium and the pre-growth medium, respectively, iii) The ratio of C NGS / C NPGS is greater than 1, where C NGS and C NPGS are the concentrations of nitrogen or a nitrogen source in the growth medium and the pre-growth medium, respectively, iv) The ratio of ΔpH GS / ΔpH PGS is less than 1, where ΔpH GS and / ΔpH PGS are the pH changes in the growth medium and the pre-growth medium, respectively, v) The ratio of Q GGS / Q GPGS is greater than 1, where Q GGS and Q GPGS are the amounts of gas, oxygen, or air introduced or bubbled into the growth medium and the pre-growth medium, respectively, vi) NSSGS / N SSPGS has a ratio less than 1, where N SSGS and N SSPGS are the number of sub - stages of the growth phase and the number of sub - stages of the pre - growth phase, respectively, where two sub - stages are separated from each other by transferring the nanoparticle - producing cells from the first sub - stage to the second sub - stage, and vii) the growth medium is supplemented with a fed - batch medium, while the pre - growth medium is not supplemented with a fed - batch medium.

6. The method according to claim 1, wherein, The pre-growth and / or growth and / or fed-batch medium contains less than per kilogram or liter of the pre-growth and / or growth and / or fed-batch medium: i) By mass or volume 5.10 -3 % or 0.5 g or 0.5 mL or 10 -8 mol or 10 -9 mol of a vitamin or chemical component selected from the group consisting of folic acid, folate, pyridoxine, pyridoxine hydrochloride, pyridoxamine, pyridoxal, riboflavin, biotin, thiamine, thiamine hydrochloride, niacin, pantothenic acid, calcium pantothenate, inositol, p-aminobenzoic acid, aminobenzoic acid, lipoic acid, all-trans retinol, retinal, alternative provitamin A-functionalized carotenoids including all-trans beta-carotene, niacin, niacinamide, niacinamide, nucleosides, cyanocobalamin, hydroxocobalamin, methylcobalamin, adenosylcobalamin, ascorbic acid, cholecalciferol, ergocalciferol, tocopherol, tocotrienol, theoquinone, menaquinone, vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin B 12 , Vitamin C, Vitamin D, Vitamin D2, Vitamin D3, Vitamin E, Vitamin K, Vitamin V i and their derivatives, where V can be any letter from A to Z, and i can be any integer from 1 to 100, ii) Six different vitamins or chemical components selected from the group consisting of: folic acid, folate, pyridoxine, pyridoxine hydrochloride, pyridoxamine, pyridoxal, riboflavin, biotin, thiamine, thiamine hydrochloride, niacin, pantothenic acid, calcium pantothenate, inositol, para-aminobenzoic acid, aminobenzoic acid, lipoic acid, all-trans retinol, retinal, alternative provitamin A-functionalized carotenoids including all-trans β-carotene, nicotinic acid, nicotinamide, nicotinamide riboside, cyanocobalamin, hydroxocobalamin, methylcobalamin, adenosylcobalamin, ascorbic acid, cholecalciferol, ergocalciferol, tocopherol, tocotrienol, menadione, phylloquinone, vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin B 12 、 vitamin C, vitamin D, vitamin D2, vitamin D3, vitamin E, vitamin K, vitamin V i and their derivatives, where V can be any letter from A to Z, and i can be any integer from 1 to 100, iii) 10 by mass or volume -2 %, or 1 g, or 1 mL, or 10 -7 mol, or 10 -8 mol of minerals or chemical components selected from the group consisting of: magnesium sulfate, sodium chloride, manganese sulfate, ferrous sulfate, ferrous sulfate heptahydrate, cobalt nitrate, calcium chloride, zinc sulfate, zinc sulfate heptahydrate, copper sulfate, copper sulfate pentahydrate, potassium alum, potassium alum dodecahydrate, sodium molybdate, sodium selenite, sodium tungstate, sodium tungstate dihydrate, nickel chloride, EDTA, MgSO4, MnSO4, NaCl, FeSO4, CoSO4, CaCl2, ZnSO4, CuSO4, KAl(SO4)2, H3BO3, Na2MoO4, NiCl2, Na2SeO3 and their derivatives, iv) 7 minerals or chemical components selected from the group consisting of: magnesium sulfate, sodium chloride, manganese sulfate, ferrous sulfate, ferrous sulfate heptahydrate, cobalt nitrate, calcium chloride, zinc sulfate, zinc sulfate heptahydrate, copper sulfate, copper sulfate pentahydrate, potassium alum, potassium alum dodecahydrate, sodium molybdate, sodium selenite, sodium tungstate, sodium tungstate dihydrate, nickel chloride, EDTA, MgSO4, MnSO4, NaCl, FeSO4, CoSO4, CaCl2, ZnSO4, CuSO4, KAl(SO4)2, H3BO3, Na2MoO4, NiCl2, Na2SeO3 and their derivatives, v) 0.005 g or 10 -8 at least one component of yeast extract of M or at least one compound derived from yeast extract, said component or compound being selected from the group of compounds consisting of: at least one protein, at least one nucleic acid, at least one functional peptide, glutathione, glucan, mannan, trehalose, flavor nucleotide, B vitamins, biotin, at least one volatile aromatic compound, calcium, phosphorus, zinc, iron, chromium, potassium, cobalt, manganese, strontium, magnesium, and their derivatives vi) 5 different components of yeast extract or different compounds derived from yeast extract, the components or compounds selected from the group consisting of: at least one protein, at least one nucleic acid, at least one functional peptide, glutathione, dextran, mannan, trehalose, flavor nucleotides, B vitamins, biotin, at least one volatile aromatic compound, calcium, phosphorus, zinc, iron, chromium, potassium, cobalt, manganese, strontium, magnesium and their derivatives, vii) 0.01 g or 10 -8 at least one peptone component of M or at least one compound derived from peptone, said component or compound being selected from the group consisting of ash, protein, sucrose, stachyose, raffinose, neutral detergent fiber, ether extract, and their derivatives viii) Different components of 5 peptones or different compounds derived from peptones, said components or compounds selected from the group consisting of: ash, protein, sucrose, stachyose, raffinose, neutral detergent fiber, ether extract, and their derivatives, ix) 0.001 grams of EDTA, x) 0.001 grams of at least one amino acid, xi) 5 different amino acids, xii) 12 different CMR, toxic or cytotoxic compounds selected from the group consisting of: manganese sulfate, cobalt nitrate, zinc sulfate, copper sulfate, potassium alum, sodium molybdate, sodium selenite, sodium tungstate, nickel chloride, and their derivatives, xiii) 5 chemical elements or heavy metals selected from the group consisting of: cadmium, lead, arsenic, mercury, cobalt, vanadium, nickel, lithium, antimony, and copper, and their derivatives, xiv) 10 -5 g is selected from the group consisting of the following chemical elements or heavy metals: cadmium, lead, arsenic, mercury, cobalt, vanadium, nickel, lithium, antimony and copper, and their derivatives, xv) 10 -5 at least one CMR, toxic or cytotoxic compound of g selected from the group consisting of: manganese sulfate, cobalt nitrate, zinc sulfate, copper sulfate, potassium alum, sodium molybdate, sodium selenite, sodium tungstate, nickel chloride and derivatives thereof, and / or xvi) 0.01 grams of peptone.

7. The method according to claim 1, wherein the concentration of at least one compound in the pre - growth and / or growth medium is C2 or the concentration is C 总量 = C1 + C2, where: C1 is the concentration of said at least one compound in the pre-growth medium and / or growth medium that is not consumed by the nanoparticle-producing cells, C2 is the concentration of said at least one compound in the pre-growth medium and / or growth medium that is consumed by the nanoparticle-producing cells, and C1 and C2 are measured or considered at the start, during, or end of the pre-growth stage and / or the growth stage.

8. The method according to claim 1, wherein the pre-growth medium, the growth medium, and / or the fed-batch medium do not comprise at least one compound at a concentration that affects the growth of the nanoparticles-producing cells and / or the production of nanoparticles, and / or wherein the pre-growth medium, the growth medium, and / or the fed-batch medium are substantially free of at least one compound, wherein the at least one compound is selected from the group consisting of: 1) Wolfe vitamins or a medium containing a total number of different components with more than half of the Wolfe vitamins, 2) one component of the Wolfe vitamins, 3) folic acid, 4) pyridoxine, 5) riboflavin, 6) biotin, 7) thiamine, 8) niacin, 9) pantothenic acid, 10) vitamin B 12, 11) aminobenzoic acid, 12) lipoic acid, 13) Wolff's minerals or a culture medium with a total number of different components containing more than half of Wolff's minerals, 14) magnesium sulfate, 15) sodium chloride, 16) manganese sulfate, 17) ferrous sulfate heptahydrate, 18) cobalt nitrate, 19) calcium chloride, 20) zinc sulfate heptahydrate, 21) copper sulfate pentahydrate, 22) potassium alum dodecahydrate, 23) sodium molybdate, 24) sodium selenite, 25) sodium tungstate dihydrate, 26) yeast extract or a culture medium with a total number of different components containing more than half of yeast extract, 27) yeast extract equivalent or a culture medium with a total number of different components containing more than half of yeast extract equivalent, 28) 1, 2 or 5 proteins derived from or contained in yeast extract, 29) 1, 2 or 5 nucleic acids derived from or contained in yeast extract, 30) 1, 2 or 5 peptides or functional peptides derived from or contained in yeast extract, 31) glutathione, 32) dextran, 33) mannan, 34) trehalose, 35) flavor nucleotides derived from or contained in yeast extract, 36) vitamin B complex, 37) biotin, 38) 1, 2 or 5 volatile aromatic compounds derived from or contained in yeast extract, 39) chromium, 40) cobalt, 41) strontium, 42) nickel chloride, 43) mineral elixir or a culture medium with a total number of different components containing more than half of mineral elixir, 44) MnSO4, 45) NaCl, 46) FeSO4, 47) CoSO4, 48) CaCl2, 49) ZnSO4, 50) CuSO4, 51) KAl(SO4)2, 52) H3BO3, 53) Na2MoO4, 54) NiCl2, 55) Na2SeO3, 56) peptone or a culture medium with a total number of different components containing more than half of peptone, 57) a component of peptone, 58) 1, 2 or 5 proteins derived from or contained in peptone, 59) sugar derived from or contained in peptone, 60) an amino acid derived from or contained in peptone, 61) ash derived from or contained in peptone, 62) a fiber derived from or contained in peptone, 63) a CMR reagent, 64) an amino acid, 65) alanine, 66) arginine, 67) asparagine, 68) aspartic acid, 69) cysteine, 70) glutamine, 71) glutamic acid, 72) glycine, 73) histidine, 74) isoleucine, 75) leucine, 76) lysine, 77) methionine, 78) phenylalanine, 79) proline, 80) serine, 81) threonine, 82) tryptophan, 83) tyrosine, 84) valine, 85) a cytotoxic or toxic compound, 86) manganese sulfate, 87) copper sulfate, 88) potassium alum, 89) sodium tungstate, 90) a heavy metal different from iron, 91) titanium, 92) vanadium, 93) manganese, 94) nickel,95) Copper, 96) Zinc, 97) Gallium, 98) Germanium, 99) Arsenic, 100) Zirconium, 101) Niobium, 102) Molybdenum, 103) Technetium, 104) Ruthenium, 105) Rhodium, 106) Palladium, 107) Silver, 108) Cadmium, 109) Indium, 110) Tin, 111) Tellurium, 112) Lutetium, 113) Hafnium, 114) Tantalum, 115) Tungsten, 116) Rhenium, 117) Osmium, 118) Iridium, 119) Platinum, 120) Gold, 121) Mercury, 122) Thallium, 123) Lead, 124) Bismuth, 125) Polonium, 126) Astatine, 127) Lanthanum, 128) Cerium, 129) Praseodymium, 130) Neodymium, 131) Promethium, 132) Samarium, 133) Europium, 134) Gadolinium, 135) Terbium, 136) Dysprosium, 137) Holmium, 138) Erbium, 139) Thulium, 140) Ytterbium, 141) Actinium, 142) Thorium, 143) Protactinium, 144) Uranium, 145) Neptunium, 146) Plutonium, 147) Americium, 148) Curium, 149) Berkelium, 150) Californium, 151) Einsteinium, 152) Fermium, 153) Niobium, 154) Radium, 155) Lawrencium, 156) Rutherfordium, 157) Dubnium, 158) Seaborgium, 159) Bohrium, 160) Hassium, 161) 162) Darmstadtium, 163) Roentgenium, 164) Copernicium, 165) Elements 113 - 118, 166) Helium, 167) Lithium, 168) Beryllium, 169) Boron, 170) Fluorine, 171) Aluminum, 172) Silicon, 173) Argon, 174) Scandium, 175) Chromium, 176) Nickel, 177) Copper, 178) Selenium, 179) Bromine, 180) Krypton, 181) Rubidium, 182) Yttrium, 183) Tin, 184) Antimony, 185) Iodine, 186) Xenon, 187) Cesium, 188) Barium, 189) Lutetium, 190) Astatine, 191) Radon, 192) Francium, 193) Mendelevium, 194) Mt, 195) Uub, 196) Uut, 197) Uuq, 198) Uup, 199) Uuh, 200) Uus, 201) Uuo, 202) Salts of Compounds 1) to 203), and 203) their derivatives.

9. The method according to claim 8, wherein the concentration of said at least one compound that affects nanoparticle - producing cell growth and / or nanoparticle production is in the pre - growth medium, growth medium, and / or fed - batch medium: greater than 1 pM or 1 μM or 1 mM or 10 -3 ng of the compound per liter of pre - growth medium, growth medium, and / or fed - batch medium, or 1 ng of the compound per liter of pre - growth medium, growth medium, and / or fed - batch medium, or 10 3 ng of the compound per liter of pre - growth medium, growth medium, and / or fed - batch medium.

10. The method according to claim 1, wherein the growth medium and / or the pre-growth medium is supplemented with a fed-batch medium, and: i) the pH value of the fed-batch medium is lower than the pH value of the pre-growth medium and / or the growth medium, and / or ii) the concentration of at least one chemical element selected from the group consisting of: a) a phosphorus source or a phosphate source, b) a potassium source, c) a magnesium source, d) an iron source, e) a vitamin source, f) a calcium source, g) KH2PO4, h) MgSO4, i) FeCl3, j) thiamine, k) CaCl2, and l) their derivatives is greater in the fed-batch medium than in the pre-growth medium and / or the growth medium.

11. The method according to claim 1, wherein: The nanoparticles are magnetosomes.

12. The method according to claim 1, wherein the pre-growth medium and / or the growth medium comprises a calcium source, a carbon source, a nitrogen source, a phosphate source or a phosphorus source, a sulfur source, an iron source, a vitamin source, and a calcium source, and: the concentration of the carbon source in the pre-growth medium and / or the growth medium is greater than the concentration of at least one compound selected from the group consisting of: a phosphate source or a phosphorus source, a sulfur source, a vitamin source, and a calcium source in the pre-growth medium and / or the growth medium, and / or the concentration of the nitrogen source in the pre-growth medium and / or the growth medium is greater than the concentration of at least one compound selected from the group consisting of: a phosphate source or a phosphorus source, a sulfur source, a vitamin source, and a calcium source in the pre-growth medium and / or the growth medium.

13. The method according to claim 1, further comprising the step of storing, amplifying, preparing, or injecting a nanoparticle-producing cell bank in the pre-growth and / or fed-batch medium and / or the growth and / or fed-batch medium, wherein the bank is stored, amplified, or prepared in a bank medium comprising at least 1% of the same compounds as the pre-growth and / or fed-batch medium and / or the growth and / or fed-batch medium.

14. A high-purity nanoparticle-producing cell obtained by the method according to claim 1, the high-purity nanoparticle-producing cell comprising more than 50% of: i) iron, based on the M FeC / M MC ratio, where M FeC is the mass of iron in the high-purity nanoparticle-producing cell, and M MC is the mass of iron and metals or metalloids other than iron in the high-purity nanoparticle-producing cell, ii) iron and at least one other metal selected from the group consisting of: sodium, magnesium, aluminum, potassium, calcium, scandium, titanium, chromium, manganese, zinc, gallium, strontium, yttrium, zirconium, niobium, molybdenum, technetium, indium, cesium, barium, lanthanum, cerium, praseodymium, neodymium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, lutetium, hafnium, rhenium, and tungsten, based on the ratio M1 / M2, where M1 is the mass of iron and at least one other metal selected from the above group in the high-purity iron oxide nanoparticles, and M2 is the mass of all metals contained in the high-purity iron oxide nanoparticles, and / or iii) iron and at least one other non-metal selected from the group consisting of: hydrogen, carbon, nitrogen, phosphorus, sulfur, fluorine, chlorine, bromine, iodine, helium, neon, argon, krypton, xenon, radon, and oxygen, based on the ratio M3 / M4, where M3 is the mass of iron and at least one other non-metal selected from the above group in the high-purity iron oxide nanoparticles, and M4 is the mass of all chemical elements contained in the high-purity iron oxide nanoparticles.

15. The high-purity nanoparticle-producing cell according to claim 14, wherein, The metal or metalloid other than iron in the high-purity nanoparticle-producing cells is selected from the group consisting of: sodium, magnesium, aluminum, potassium, calcium, scandium, titanium, chromium, manganese, zinc, gallium, strontium, yttrium, zirconium, niobium, molybdenum, technetium, indium, cesium, barium, lanthanum, cerium, praseodymium, neodymium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, lutetium, hafnium, rhenium, and tungsten, and their derivatives, at least 5 different metals or metalloids.

16. A composition comprising the high-purity nanoparticle-producing cell according to claim 14.

17. A high-purity iron oxide-based nanoparticle obtained by the method according to claim 1, the high-purity iron oxide-based nanoparticle comprising more than 93% of: i) iron, based on the ratio of M FeN / M MN where M FeN is the mass of iron in the high-purity iron oxide nanoparticles, and M MN is the mass of iron and metals or metalloids other than iron in the high-purity iron oxide nanoparticles, ii) iron and at least one other metal selected from the group consisting of: sodium, magnesium, aluminum, potassium, calcium, scandium, titanium, chromium, manganese, zinc, gallium, strontium, yttrium, zirconium, niobium, molybdenum, technetium, indium, cesium, barium, lanthanum, cerium, praseodymium, neodymium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, lutetium, hafnium, rhenium, and tungsten, based on the ratio M1 / M2, where M1 is the mass of iron and at least one other metal selected from the above group in the high-purity iron oxide nanoparticles, and M2 is the mass of all metals contained in the high-purity iron oxide nanoparticles, and / or iii) iron and at least one other non-metal selected from the group consisting of: hydrogen, carbon, nitrogen, phosphorus, sulfur, fluorine, chlorine, bromine, iodine, helium, neon, argon, krypton, xenon, radon, and oxygen, based on the ratio M3 / M4, where M3 is the mass of iron and at least one other non-metal selected from the above group in the high-purity iron oxide nanoparticles, and M4 is the mass of all non-metal elements contained in the high-purity iron oxide nanoparticles.

18. The iron oxide-based nanoparticles of high purity according to claim 17, wherein the metal or metalloid other than iron in the high-purity iron oxide nanoparticles is selected from the group consisting of: sodium, magnesium, aluminum, potassium, calcium, scandium, titanium, chromium, manganese, zinc, gallium, strontium, yttrium, zirconium, niobium, molybdenum, technetium, indium, cesium, barium, lanthanum, cerium, praseodymium, neodymium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, lutetium, hafnium, rhenium, tungsten and their derivatives, at least 5 different metals or metalloids.

19. The iron oxide-based nanoparticles of high purity according to claim 17, wherein the iron oxide-based nanoparticles of high purity are magnetosomes.

20. A composition comprising the iron oxide-based nanoparticles of high purity according to claim 19.

21. A composition comprising cells produced from high-purity nanoparticles obtained by the method according to claim 1 and high-purity iron oxide nanoparticles, wherein: The high-purity nanoparticle-producing cells contain more than 50% of: i) Iron, based on the ratio of M FeC / M MC where M FeC is the mass of iron in the high-purity nanoparticle-producing cells, and M MC is the mass of iron and metals or metalloids other than iron in the high-purity nanoparticle-producing cells ii) iron and at least one other metal selected from the group consisting of: sodium, magnesium, aluminum, potassium, calcium, scandium, titanium, chromium, manganese, zinc, gallium, strontium, yttrium, zirconium, niobium, molybdenum, technetium, indium, cesium, barium, lanthanum, cerium, praseodymium, neodymium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, lutetium, hafnium, rhenium, and tungsten, based on the ratio M1 / M2, where M1 is the mass of iron and at least one other metal selected from the above group in the high-purity iron oxide nanoparticles, and M2 is the mass of all metals contained in the high-purity iron oxide nanoparticles, and / or iii) iron and at least one other non-metal selected from the group consisting of: hydrogen, carbon, nitrogen, phosphorus, sulfur, fluorine, chlorine, bromine, iodine, helium, neon, argon, krypton, xenon, radon, and oxygen, based on the ratio M3 / M4, where M3 is the mass of iron and at least one other non-metal selected from the above group in the high-purity iron oxide nanoparticles, and M4 is the mass of all non-metal elements contained in the high-purity iron oxide nanoparticles, and The high-purity iron oxide nanoparticle-based contains more than 93% of: i) Iron, based on the ratio of M FeN / M MN where M FeN is the mass of iron in the high-purity iron oxide nanoparticles, and M MN is the mass of iron and metals or metalloids other than iron in the high-purity iron oxide nanoparticles. ii) iron and at least one other metal selected from the group consisting of: sodium, magnesium, aluminum, potassium, calcium, scandium, titanium, chromium, manganese, zinc, gallium, strontium, yttrium, zirconium, niobium, molybdenum, technetium, indium, cesium, barium, lanthanum, cerium, praseodymium, neodymium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, lutetium, hafnium, rhenium, and tungsten, based on the ratio M1 / M2, where M1 is the mass of iron and at least one other metal selected from the above group in the high-purity iron oxide nanoparticles, and M2 is the mass of all metals contained in the high-purity iron oxide nanoparticles, and / or iii) iron and at least one other non-metal selected from the group consisting of: hydrogen, carbon, nitrogen, phosphorus, sulfur, fluorine, chlorine, bromine, iodine, helium, neon, argon, krypton, xenon, radon, and oxygen, based on the ratio M3 / M4, where M3 is the mass of iron and at least one other non-metal selected from the above group in the high-purity iron oxide nanoparticles, and M4 is the mass of all non-metal elements contained in the high-purity iron oxide nanoparticles.

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