title: [Expression and activity determination of TNFR domain of osteoprotegerin in E.coli and corresponding antibody preparation].
abstract: Osteoprotegerin (OPG) plays an important role in the regulation of bone resorption and remodeling. The TNFR domain of OPG, which is involved in the inhibition of formation and activity of osteoclasts, was amplified by PCR and inserted into multiple cloning site of PET-28a. The recombinant plasmid was transferred into E.coli BL21 to express recombinant protein. It was found that expressed product existed in the form of inclusion body. The inclusion body was solubilized, renatured and purified by affinity chromatography. Polyclonal antibodies with high specificity were obtained from the serum of rabbit immunized with purified recombinant protein. Mice were used to determine the hypocalcemic effect of the recombinant protein. Results showed that the recombinant protein expressed in E.coli had the proper bioactivity.
authors.affiliation: Hangzhou Institute of Calibration and Testing for Quality and Technical Supervision, Hangzhou 310004, China. xiao9988@126.com
authors.forename: Hai-Long
authors.initials: HL
authors.lastname: Xiao
authors.forename: Ai-Xia
authors.initials: AX
authors.lastname: Ren
authors.forename: Yao-Zhou
authors.initials: YZ
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D002118:Calcium / Q000097:blood; D004587:Electrophoresis, Agar Gel / Q000379:methods; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D053244:Osteoprotegerin / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011817:Rabbits; D011897:Random Allocation; D018124:Receptors, Tumor Necrosis Factor / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D014162:Transfection
chemical_list: D000906:Antibodies; D053244:Osteoprotegerin; D018124:Receptors, Tumor Necrosis Factor; D011994:Recombinant Proteins; D002118:Calcium