Reagent capable of preventing and eliminating pollution of mammalian cells

Abstract

The invention provides a reagent capable of preventing and eliminating pollution of mammalian cells, and belongs to the field of microbial pollution control in mammalian cell culture. A polypeptide having a sequence of SEQ ID NO:1 can be used for preventing and eliminating pollution of the mammalian cells; the reagent is composed of the polypeptide having the sequence of SEQ ID NO:1 and amino acid and does not contain any antibiotic component. The product is prepared in accordance with the concentration required for prevention and elimination of pollution, microbial pollution in the mammalian cells can be effectively prevented and eliminated, and the defects of use restriction of antibiotic components in a conventional product and longer cell treatment period are overcome; and the product is good in stability and easy to preserve.

Description

technical field [0001] The present invention relates to the field of microbial contamination control in mammalian cell culture. Background technique [0002] Cell culture technology is widely used in scientific research, teaching, clinical experimental research, research and development and production of blood products, antibodies, vaccines and other biological products. The quality of cells directly affects the results of scientific research and the safety and quality of biological products. In the process of cell culture, the threat of microbial contamination is ubiquitous. Potential contamination mainly comes from: potential sources of contamination carried by cells themselves, contamination of raw and auxiliary materials, cross-contamination between cells, and operational contamination. [0003] In mammalian cell culture, the characteristics of bacterial contamination are obvious, and the culture medium can turn yellow in a short period of time and appear obviously turbi...

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Problems solved by technology

[0007] As can be seen from the above table, most of the main antimicrobial agents on the market are antibiotic preparations. Although antibiotics are convenient for preventing and removing cell contamination, there are also many disadvantages: 1. The use of antibiotics will cause damage to cells in varying degrees (SPLangdon, Cell culture contamination: an overview[J]. Methods Mol Med, 2004,88:309e17.), and is dependent (Wang Hong et al., Evaluation of Rescue Methods for Contamination of Precious Adherent Cells in Cell Culture, Journal of Capital Medical University, 2011, 3 (1): 129-135.); 2. The problem of drug dependence, the mechanism of action of traditional antibiotics is mostly through hindering the synthesis of bacterial cell walls, inhibiting protein synthesis, affecting the metabolism of nucleic acid and folic acid, etc., and microorganisms can easily pass through Genetic mutations to alter metabolic pathways that lead to resistance to antibiotics

3. Mycoplasma has no cell wall, and antibiotics that usually act on cell wall biosynthesis, such as lactams and vancomycin, are completely insensitive to it; 4. The 2015 edition of the Pharmacopoeia has clear restrictions on the use of antibiotics in biological products: unless otherwise stated Unless otherwise specified, penicillin or other β-lactam antibiotics shall not be used; in the production process, the use of antibiotics shall be avoided as much as possible, and when necessary, the use of no more than one type shall be used; in the inspection of finished products, the residual amount limit shall be tested and specified; 5 . It takes a long time to treat the contaminated cells with antibiotics, usually 1-2 weeks. The physiological properties of the cells after treatment may change, which will affect the test results or the quality of the final biological products.

Although there are many types of antimicrobial peptides, few are used in production practice. The reasons are as follows: 1. Antimicrobial peptide molecules are small, difficult to separate and purify, and are easily degraded by proteases; 2. Most antimicrobial peptides have the characteristics of attacking microbial cell membranes. It is toxic to eukaryotic cells (such as human cells); 3. Antimicrobial peptides have different properties, and a lot of basic research work must be done for specific intended uses, including comprehensive pharmacological and toxicological tests, to prove sufficient safety and Effectiveness can continue; 4. Compatibility of components in antimicrobial peptide preparations, antimicrobial peptides are biologically active substances, many factors will reduce the activity of antimicrobial peptides, such as high concentrations of monovalent and divalent cations, polyanions ( polyanion), serum, apolipoprotein A-I (apolipoprotein A-I ) and protease, etc.

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