A method for rapid preparation of multiple anti-antibiotic hybridoma monoclonal antibodies

Abstract

The invention discloses a novel method for quickly and efficiently preparing a plurality of antibiotic-resistant hybridoma monoclonal antibodies in a time-saving manner. The method comprises the following concrete steps: (1) covalently linking different antibiotics to a same protein carrier, and immunizing a mouse with a cross-linked antibiotic protein conjugate; (2) separating spleen cells of the mouse, fusing the spleen cells of the mouse with myeloma cells SP2 / 0 of the mouse, and performing selective culture; (3) screening out positive hybridoma cell strains for producing different antibiotic-resistant monoclonal antibodies, and storing respectively; (4) preparing single antibiotic-resistant monoclonal antibody from the different hybridoma cell strains in the step (3). According to the method, the monoclonal antibodies resistant to different antibiotics can be obtained through one-time fusing at the same time, so that the preparation time of the monoclonal antibodies is greatly shortened and the preparation cost of the monoclonal antibodies is greatly reduced.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an antibiotic protein conjugate and a novel method for preparing various anti-antibiotic hybridoma monoclonal antibodies rapidly, efficiently and time-savingly using the conjugate. Background technique [0002] Milk is a nutritional product suitable for all ages, but with the widespread use of antibiotics in dairy farming, the issue of antibiotic residues in milk has drawn increasing attention from the international community. In 1991, the U.S. Food and Drug Administration (FDA) tested 234 fresh milk samples and found only one sample had residues of the antibiotic sulfamethazine. In the first half year of 1992, among the 197 fresh milk samples of the U.S. Food and Drug Administration (FDA), 4 samples had antibiotic sulfonamide residues. European and American countries stipulate that milk and its dairy products that exceed the allowable standard are all discarded and must...

AI Technical Summary

Problems solved by technology

However, due to the small-scale and scattered characteristics of milk production in my country, it is difficult to implement the "Industrial Standards for Pollution-free Food Fresh Milk Industry", resulting in the problem of antibiotic residues in milk and dairy products in my country. The problem of milk safety is still a huge problem. question

Although the technology of immunizing mice with multiple antigens to produce multiple antibodies has been established, for the preparation of anti-antibiotic monoclonal antibodies, immunizing mice with multiple antibiotic carrier protein conjugates at the same time, due to the excessive amount of carrier protein Large, it is easy to form immune tolerance of mice to carrier protein, and the immunosuppressive effect of carrier protein inhibits the immune response of mice to antibiotics, thus leading to a decrease in the titer of anti-antibiotic antibodies, and it is difficult to obtain anti-antibiotic monoclonal antibody hybridization tumor cell line

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