Poly-clonal antibody for specifically identifying protein A, as well as preparation method and application thereof
A polyclonal antibody and protein recognition technology, which is applied in the field of polyclonal antibody and its preparation, can solve problems affecting immune detection, etc., and achieve the effects of easy breeding, high expression yield, and guaranteed expression
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Embodiment 1
[0026] Example 1: Preparation of polyclonal antibody specifically recognizing protein A
[0027] 1. Epitope Analysis
[0028] DNAStar software was used for epitope analysis, and the analysis conclusion showed that the antigenicity of the sequence shown in SEQ ID No.1 was good, and the antigenic determinants were mainly concentrated in the N-terminal 19 amino acids and the C-terminal 45 amino acids. According to the results of the antigenic determinant analysis, Consider prokaryotic expression of SEQID No.1, and design specific primers based on the sequence.
[0029] 2. Construction of expression vector
[0030] 1. Acquisition of protein A target fragment
[0031] In Beijing Tianyi Huiyuan Biotechnology Co., Ltd., the DNA sequence used for the construction of the prokaryotic expression vector was synthesized by the method of whole gene synthesis, that is, the nucleotide sequence shown in SEQ ID No.1 was synthesized.
[0032] 2. PCR amplification
[0033] With the nucleotide...
Embodiment 2
[0051] Embodiment 2: ELISA method is used to quantitatively detect the content of protein A in the test sample
[0052] Protein A affinity chromatography is widely used in the purification of antibodies. In this method, Protein A is bound to the purification medium by covalent coupling. The shedding of Protein A during purification is a major problem in drug research. The polyclonal antibody of the present invention that specifically recognizes protein A is applied to the residual level of protein A in the quantitative detection test sample, and it is also convenient for the protein A purification medium manufacturer to monitor the shedding characteristics of protein A under specific conditions.
[0053] In this example, the principle of double-antibody sandwich enzyme-linked immunoassay (sandwich ELISA) was used to detect the residue of Protein A, and the microwell plate was coated with a monoclonal antibody that specifically recognized protein A to form a solid-phase antibody...
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