Preparation of murine monoclonal antibody

   Murine monoclonal antibody is the most common and classic monoclonal antibody. Hybridoma cells are obtained by cell fusion of B lymphocytes treated with specific antigen and myeloma cells. Positive clone strains are obtained after Hat Medium screening and ELISA detection of positive rate. Finally, cells are cultured or injected into the abdominal cavity of BALB / c mice and cultured with ascites, Monoclonal antibodies can be obtained after ascites collection and purification.

Preparation process of murine monoclonal antibody：

1. Antigen preparation: prokaryotic expression of proteantigen, or design and synthesize polypeptide coupled with KLH. Customers who choose prokaryotic expression proteantigen can provide purified antigen (purity > 85%) by themselves, Topbiotech can also purify protein for you, from constructing prokaryotic expression plasmid to obtaining purified protein. Customers who select coupled polypeptide, the antigen is prepared and synthesized by Topbiotech.

2. Antigen immunization: four BALB/c mice are immunized, and the immunization is strengthened through standard procedures. The tail blood is collected for ELISA detection and the acceptance standard is serum ELISA > 10000.

3. Cell fusion and screening: BALB/c mice with the best immune response will be chosen and its spleen cells will be fused with myeloma cells. The hybridoma cells will be screened through hat selective medium.

4. Hybridoma cell screening: hybridoma cells are screened by limited dilution analysis (LDA), 2-5 positive mother cells are subcloned, and one subclone is selected for subtype identification.

5. Mass preparation of monoclonal antibodies: a positive clone is intraperitoneally injected into 5 BALB/c mice for ascites production.

6. Monoclonal antibody purification: the antibody is purified by protein A/G, and the valence of antibody is detected by ELISA.

Our advantages：

1. In addition to providing conventional monoclonal antibody preparation of protein and polypeptide antigens, Topbiotech is also good at monoclonal antibody preparation of small molecule antigens, coupling and immunization of pesticide, small molecule and heavy metal antigens.

2. High quality and diversity of antigens: Topbiotech provides the recombinant protein expressed by prokaryotic, with purity ≥ 95%, concentration up to 5mg/mL, good solubility and simulating the state of natural protein. Topbiotech uses proprietary antigen epitope prediction software to screen and design polypeptide sequences, and the antigen immune effect is good.

3. Professional antibody screening and purification technology: Topbiotech has protein A/G antibody purification technology and antigen-specific affinity purification technology. You choose!

4. Engaged in antibody production and customized services, we are professional! Topbiotech promises that the ELISA titer of monoclonal antibody will reach more than 1:64000. The monoclonal antibody service for any host and any protein antigen (purity > 85%) ensures that the test result of Western blot is positive.

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Notes to customers:

1. For the antigen provided by the customer, the concentration should be > 85%, the molecular weight is > 10kDa, and the concentration is preferably between 1-5mg/mL.

2. The amount of antigen that the customer needs to provide is determined by the immune scheme. If antigen-specific purification of antibody is required, more antigen is required.

3. The customer is responsible for the materials and information provided. In case of experimental delay or economic loss caused by inaccurate materials and information provided by the customer, the customer should bear all the expenses.