Screening and prokaryotic expression purification of nano antibody against Leucine aminopeptidase of Echinococcus multilocularis
Objective To screen an anti-leucine aminopeptidase of Echinococcus multilocularis(Em-LAP)nano antibody so as to construct a prokaryotic expression system of the antibody.Therefo re,the nano antibody pro-tein was expressed and purified to obtain the nano antibody against leucine aminopeptidase of Echinococcus multi-locularis with high specificity.Methods The phage library display technology was utilized for screening a nano an-tibody against leucine aminopeptidase.The affinity of nano antibody to the target protein was verified by phage-ELISA.After sequencing the antibody with high affinity,the target sequence was concatenated and the codon was optimized.After synthesis,it was inserted into the expression vector pET30a for enzyme digestion identification.The recombinant plasmid was transferred into Escherichia coli BL21(DE3)strain,and isopropyl-β-D-thiogalactopy-ranoside(IPTG)was used to induce the expression of the target protein.The recombinant target protein LAPNb was purified by metal-chelating affinity chromatography.Results The anti-Em-LAP nano antibody was screened,and the recombinant pET30a-LAPNb plasmid was constructed.A protein with a size of about 28 KD was obtained.Con-clusion The study screened and obtained anti-Em-LAP nano antibody,and constructed a LAPNb prokaryotic ex-pression system.The protein LAPNb was expressed and purified.
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