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Preparation Of Monoclonal Antibody Against Capsaicin And Development Of Colloidal Gold Immunochromatography Strip

Posted on:2022-10-07Degree:MasterType:Thesis
Country:ChinaCandidate:H M KangFull Text:PDF
GTID:2480306734967229Subject:Engineering Bioengineering
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Objective:To prepare capsaicin monoclonal antibody with strong specificity and high inhibition rate for preparation of colloidal gold lateral immunochromatography strip and for rapid detection of Capsaicine in vegetable oil in situ.Method:The capsaicin hapten was chemically coupled with BSA and OVA,respectively,to obtain capsaicin-BSA and capsaicin-OVA complete antigens.BALB/c mice were immunized with capsaicin-BSA as immunogens,and spleen cells of immunized mice were fused with SP2/0 myeloma cells.Hybridioma cell lines secreting stable monoclonal antibodies against capsaicin were screened by limited dilution method.Preparation and purification of ascites type monoclonal antibody,and identification of specificity,IC50,concentration,purity,titer and other characteristics.Monoclonal antibody against capsaicin was labeled on Au nanopaticles,and capsaicin-OVA-complete antigen was coated on the test line.Based on the competitive lateral immunochromatography strategy,a competitive lateral immunochromatography strip for capsaicin was established and its performance and application were evaluated.Results:Three hybridoma cell lines with specific monoclonal antibodies against capsaicin were screened.Based on 5E12 monoclonal antibody with the best specificity and affinity,capsaicin colloidal gold lateral immunochromatography test strip was established.The sensitivity of the method was 10 ng/m L,the reaction time was 10min,the specificity was good,and it could be used to detect the 10-times concentrated extract of edible oil.The positive and negative coincidence rates were 100%and88.8%respectively,which could meet the requirements of rapid detection of"illegal cooking oil"in the field.Conclusion:Monoclonal antibody against capsaicin was successfully prepared,and colloidal gold lateral immunochromatography strip was developed.The method has high sensitivity,high specificity,simple operation and rapid reaction,which meets the requirements of rapid identification and detection of"illegal cooking oil"food safety in the field.
Keywords/Search Tags:Capsaicin, cooking oil, AuNPs, immunochromatography, food safety
PDF Full Text Request
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