Lyophilized formulation of a monoclonal antibody against transthyretin
A monoclonal antibody, freeze-dried preparation technology, applied in the direction of anti-animal/human immunoglobulin, antibody, freeze-dried delivery, etc., can solve problems such as cytotoxicity
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Embodiment 1
[0278] Example 1. Low-concentration pre-formulation test
[0279] Preformulation development studies were performed on the humanized 14G8 antibody with the mature heavy chain sequence of SEQ ID NO:82 (except that the C-terminal lysine may be absent) and the mature light chain sequence of SEQ ID NO:86. Pre-formulation tests were performed with the humanized 14G8 antibody in 20 formulations containing combinations of various buffers in the pH range of about 4.5 to about 7.0, with and without certain sugars, surfactants and other excipient.
[0280] By including visual inspection, dynamic light scattering (DLS), microfluidic imaging (MFI), differential scanning fluorescence (DSF), differential scanning calorimetry (DSC) and high performance size exclusion chromatography (HP-SEC) method for testing the properties of formulations.
[0281] The buffers tested were citrate, histidine, succinate and phosphate at concentrations ranging from about 10 mM to about 20 mM. The sugars tes...
Embodiment 2
[0293] Embodiment 2: Formulation screening of 50mg / ml
[0294] Formulations F21-F31 listed in Table 4 were tested at a concentration of 50 mg / ml antibody based on the results of the pre-formulation screen.
[0295] Table 4: List of formulations tested at 50 mg / ml
[0296] Table 4
[0297]
[0298] Prepared (T-liquid), lyophilized (T0), stored in liquid state at 25°C for two weeks and stored in lyophilized form at 2-8°C, 25°C and 40°C for one month and three months as appropriate The samples were subjected to visual inspection, glass transition temperature, MFI, HP-SEC and cIEF tests after 1 month. In each of these tests, the results of the formulations were assigned "poor", "acceptable", "good" and "very good" values. Acceptable formulations have 4000-6000 particles ≥10 μm / ml (F27), greater than 94.0% monomers and less than 3% high molecular weight species after storage at 40°C / 75% relative humidity (r.h.) for three months (F22-F27 and F30), and greater than 74% of th...
Embodiment 3
[0306] Example 3: Preparation and Characterization of Formulation F25
[0307] Sample Preparation : The liquid formulation before lyophilization was 61 mg / ml humanized 14G8, 20 mM histidine, 240 mM sucrose and 0.04% poloxamer. After lyophilization, the formulation was reconstituted to an antibody concentration of approximately 50 mg / ml.
[0308] Thaw the frozen stock as described above in a water bath at 20 °C. After thawing, the two batches of material were mixed and the protein concentration was determined. An aliquot of the second batch was re-frozen to -80 °C until further use. Sample concentrations were adjusted by dilution with formulation buffer to achieve a target concentration of 50 mg / ml. Surfactants were added by adding a small aliquot of the concentrated stock solution in water directly to the formulation. Then, the samples were filtered through a 0.22-μm PVDF membrane filter under a laminar flow hood. Finally, the filtered formulation was filled into pre-cl...
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