Expression And Purification Of T7 Bacteriophage Capsid Protein P11 And Preparation Of Monoclonal Antibody Against P11

Bacteriophage T7 is a lysogeny phage capable of infecting most strains of Escherichia coli, a double standed DNA of genome molecular is 39 936 bp. The capside protein of T7 include major head protein P10A, head assemble protein P10B, neck protein P8, Tail protein P11, P12 and Tail fibers protein P17. T7 phage display system is easy to use and has the capacity to display peptides up to about 1200 amino acids, including stability of insert gene, varity of elution conditions and short of growth cycle. T7 phage protein chip is fixed the bacteriophage T7 which expressed and displayed the exogenous proteins on the chip, the chip could be used for the detection the exogenous proteins. P11 gene was inserted into plasmid pET-28a(+) to construct recombinant plasmid pET-28a(+)/P11. Recombinant P11 protein was expressed under the introduction of 0.1mol/L IPTG and well purified by nickel ion affinity chromatogagh method. SDS-PAGE analysis showed that the molecular weight of the expressed protein was approximately 27 kDa. The Balb/c mice were immunized with the purified protein P11 mixed adjuvant. The monoclonal antibodys (2P11) were identified with indirect immunized fluresent assay and Western blot assay. Indirect ELISA was used to detect the Ig isotype.We acquired the recombinant P11 protein and prepared a strain of hybridoma cell secreting the MAbs against P11 protein, the MAbs are useful material in vitro diagnosis.