title: Characterization of recombinant soluble human transforming growth factor-beta receptor type II (rhTGF-beta sRII).
abstract: Recombinant human transforming growth factor soluble receptor Type II (rhTGF-beta sRII) corresponding to the 159 amino acid extracellular domain of hTGF-beta RII has been expressed in insect cells using the baculovirus expression system or in a mouse myeloma cell line. N-terminal sequence analysis of the purified protein revealed the removal of the 23 amino acid signal peptide. In SDS-PAGE, the rhTGF-beta sRII resolves into multiple bands due to N-linked glycosylation. Recombinant hTGF-beta sRII is a TGF-beta antagonist and will inhibit the biological activities of TGF-beta 1, TGF-beta 3, and TGF-beta 5 on TGF-beta-responsive cell lines, such as murine HT-2 or human TF-1 with an ED50 of approximately 0.3 micrograms/mL. However, hTGF-beta sRII does not inhibit TGF-beta 2 bioactivities in these cell lines, suggesting that hTGF-beta RII has low affinity for TGF-beta 2. Polyclonal antibodies to hTGF-beta sRII have been produced in goats and purified on Protein-G affinity columns. This antibody can inhibit TGF-beta 1,2,3,5-dependent bioactivities on human cell lines such as TF-1. Additionally, this antibody has species cross-reactivity and will also inhibit TGF-beta-dependent bioactivities on murine cells.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: R&D Systems, Minneapolis, MN 55413, USA.
authors.forename: M L
authors.initials: ML
authors.lastname: Tsang
authors.forename: L
authors.initials: L
authors.lastname: Zhou
authors.forename: B L
authors.initials: BL
authors.lastname: Zheng
authors.forename: J
authors.initials: J
authors.lastname: Wenker
authors.forename: G
authors.initials: G
authors.lastname: Fransen
authors.forename: J
authors.initials: J
authors.lastname: Humphrey
authors.forename: J M
authors.initials: JM
authors.lastname: Smith
authors.forename: M
authors.initials: M
authors.lastname: O'Connor-McCourt
authors.forename: R
authors.initials: R
authors.lastname: Lucas
authors.forename: J A
authors.initials: JA
authors.lastname: Weatherbee
mesh_terms: D000818:Animals; D000918:Antibody Specificity; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors; D006801:Humans; D007313:Insecta; D051379:Mice; D008907:Mink; D008969:Molecular Sequence Data; D018125:Receptors, Transforming Growth Factor beta / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D012680:Sensitivity and Specificity; D012995:Solubility; D016212:Transforming Growth Factor beta / Q000037:antagonists & inhibitors
chemical_list: D018125:Receptors, Transforming Growth Factor beta; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta