Metformin monoclonal antibody hybridoma cell line and application thereof
A technology of hybridoma cell lines and monoclonal antibodies, which is applied in the preparation methods of animal/human proteins and peptides, serum albumin, etc., can solve the problems of high detection limit, complexity, and low sample purity requirements, and achieve good detection sensitivity , good specific effect
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[0023] Example 1 Preparation of hybridoma cell line Tcf 1A6
[0024] (1) Synthesis of complete antigen: Weigh 1.8 mg of MET derivative 4cpMET (the molar ratio of 4cpMET to bovine serum albumin (BSA) is 30:1), dissolve it in 300 μL N,N-dimethylformamide (DMF) , stirred and reacted for 10 minutes, after fully dissolving, 1.3 mg of N-hydroxysuccinimide (NHS) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) were successively added 2.1 mg, activated at room temperature for 4-6 hours, as solution A. Take 6mg of BSA, dissolve it with 2mL 0.01M carbonate buffer solution (CB, pH=9.0) (referred to as solution B), then slowly add solution A to solution B drop by drop, and perform coupling reaction at room temperature overnight; then use 0.01 M PBS solution was dialyzed to remove the unreacted small molecule hapten to obtain the complete antigen 4cpMET-BSA, which was identified by ultraviolet absorption scanning method;
[0025] (2) Animal immunization: After mixing...
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