Preparation of 5-methyl morpholine-3-amino-2-oxazolidinone derivative hapten, antigen and antibody
A technology of methylmorpholine and oxazolidinone, applied in the field of immunology, can solve problems such as unestablished, and achieve the effect of easy identification, high potency and stable results
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Embodiment 1
[0037] Example 1 Preparation method of 5-methylmorpholine-3-amino-2-oxazolidinone derivative hapten MCPAMOZ
[0038] Add 1.15g of p-carboxybenzaldehyde into a 50ml round bottom flask, slowly add methanol until the p-carboxybenzaldehyde is completely dissolved, add 0.5g of 5-methylmorpholine-3-amino-2-oxazolidinone while stirring, and stir at room temperature overnight. Reaction finishes filtering, washes twice with water, washes twice with methanol, is dried, and obtains 0.80g white powder, and its structural formula is as shown in (I). ESI-MS (negative) m / z: 332 (M-H) - . 1 H NMR (400MHz, d 5 -Pyridine, TMS): δ8.44 (d, J = 8.3, 2H); 7.99 (d, J = 8.3, 2H) 7.94 (s, 1H); 4.96 (qd, J 1 =6.2,J 2 =6.2,J 3 =6.3,J 4 = 8.4, 1H); 4.09 (t, J = 8.9Hz, 1H); 3.78 (dd, J1 = 6.7, J2 = 8.8, 1H); 3.69-3.64 (m, 4H); 2.67 (dd, J 1 =6.2,J 2 = 13.3, 1H); 2.61 (dd, J 1 =5.7,J 2 =13.4); 2.51-2.46 (m, 4H)
[0039]
[0040] (I)
Embodiment 2
[0041] Example 2 Preparation method of 5-methylmorpholine-3-amino-2-oxazolidinone derivative hapten CEPAMOZ
[0042] Add 1.06 g of 2-(4-formylphenoxy)acetic acid into the round bottom flask, slowly add methanol until the p-carboxybenzaldehyde is completely dissolved, add 0.5 g of 5-methylmorpholine-3-amino-2-oxadiene while stirring Oxazolidinone, stirred overnight at room temperature. After the reaction was completed, filtered, washed twice with water and twice with methanol, and dried to obtain 0.93 g of white powder, the structural formula of which was shown in formula (II). ESI-MS (negative) m / z: 362 (M-H) - . 1 H NMR (600MHz, d 5-Pyridine, TMS): δ7.89-7.87 (m.3H); 7.22 (d, J=8.8, 2H); 5.02 (s, 2H); 4.88 (qd, J 1 =6.31, J2=6.31, J3=6.35, J4=8.5, 1H); 4.01(t, J=8.8Hz, 1H); 3.70(dd, J1=6.8, J2=8.7, 1H); 3.67-3.65(m , 4H); 2.63(dd, J1=6.2, J2=13.3, 1H); 2.56(dd, J1=5.6, J2=13.4); 2.51-2.44(m, 4H)
[0043]
Embodiment 3
[0044] The preparation method of embodiment 3 immunogen CEPAMOZ-BSA
[0045] Dissolve 0.1mmol of the hapten CEPAMOZ in 2ml of DMF, stir and add 27.5mg of DCC and 14.4mg of NHS, and react overnight with magnetic stirring at 4°C. After centrifugation, the supernatant is liquid A; weigh 140mg of BSA and dissolve in 10ml with a concentration of 0.1mol / L Add 1ml of DMF to the PBS (pH8.0), stir and dissolve to prepare liquid B; under magnetic stirring, liquid A is gradually dropped into liquid B, and react at 4°C for 12 hours. After centrifugation, take the supernatant, dialyze with normal saline at 4°C for 3 days, change the dialysate 3 times a day, and distribute the whole antigen in 0.5ml at a concentration of 1mg / ml
[0046] Store in a centrifuge tube and freeze at -20°C for immunization. The structure of the prepared 5-methylmorpholine-3-amino-2-oxazolidinone derivative immunogen CEPAMOZ-BSA is shown in formula (V):
[0047]
[0048] Immunogen identification: The carrier p...
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