Preparation method of bombyx mori silk fibroin monoclonal antibody

Abstract

The invention relates to the field of archaeological detection and discloses a preparation method of a bombyx mori silk fibroin monoclonal antibody. The preparation method comprises the following steps: extracting bombyx mori silk fibroin by adopting an isooctane, n-octyl alcohol, bi(2-ethylhexyl) sodium sulfosuccinate and polyvinyl pyridine quaternary ammonium salt mixed system, injecting the bombyx mori silk fibroin serving as a complete antigen into the bodies of rabbits, selecting the rabbits with high immunizing potency, fusing splenic lymphocytes and myeloma cells, cultivating the obtained hybridoma cells, selecting strains with positive secretory antibodies, high secretion ability and good cell growth state by an indirect ELISA method, cloning by a limited dilution method until the antibody secretion positive rate of hybridoma cell growth is 100 percent, performing big turn production on the screened cells, and purifying the obtained monoclonal antibody by a chromatographic column. The antibody prepared by the method has high specificity, and is simple and rapid in operation during the application process and high in detection accuracy.

Description

Technical field [0001] The invention relates to the field of archaeological detection, in particular to a method for preparing a mulberry silk fibroin monoclonal antibody. Background technique [0002] Silk is a kind of natural polymer protein fiber, mainly divided into two categories: tussah silk and mulberry silk. In view of the fact that the origin of mulberry silk has been shrouded in the mist, a more scientific method is urgently needed to identify the unearthed silk. The conventional detection methods of silk mainly include Fourier infrared spectroscopy, Raman spectroscopy, X-ray diffraction, etc. However, long-term use of silk in the environment of tombs or ruins will be affected by various factors, resulting in protein degradation and macromolecular chain breakage. It is difficult to detect the presence of silk fibroin by some traditional methods. Therefore, how to use the advanced methods of natural science to establish a silk fabric micro-mark detection technology sy...

AI Technical Summary

Problems solved by technology

At present, polyclonal antibodies to silk fibroin protein have appeared. The specificity of antibodies depends on the determinants of antigen molecules. Antigen molecules have many antigen determinants. Therefore, the antibodies produced by immunized animals are mixtures of various antibodies. It is very difficult to separate the antibodies, and polyclonal antibodies have problems such as low purity, uneven physical and chemical properties, and weak binding specificity to antigens.