Comparative studies on the interaction of proteins with a polydimethylsiloxane elastomer. II. The comparative antigenicity of primary and secondarily adsorbed IgG1 and IgG2a and their non-adsorbed counterparts.
The antigenicity of bovine IgG1 and IgG2a adsorbed on a polydimethysiloxane (PEP) elastomer, on a widely used polystyrene (Imm 2, Dynatech) or immobilized as biotinylated proteins to streptavidin covalently bound to polystyrene (SA-PS) was compared using various monoclonal (mAbs) and polyclonal antibodies (pAb) to bovine IgG. The IgGs were either adsorbed as native proteins or pre-denatured with 6M Guanidine-HCl (Gu-HCl) or 6 M Gu-HCl/0.1% 2-mercaptoethanol. In special situations, bovine and human IgG was immobilized by secondary adsorption to an albumin monolayer adsorbed on either PEP or Imm 2. Results indicate that pre-denaturation of IgGs with 6 M Gu-HCl/2-mercaptoethanol destroys all antigenicity whereas those IgGs pretreated with 6 M-GuHCl are indistinguishable in their antigenicity from the IgGs adsorbed to either PEP or Imm 2 without such treatment. When immobilized on SA-PS, Gu-HCl-treated IgGs were significantly less detectable, especially when tested using mAbs. In general, IgGs adsorbed on PEP or Imm 2 were less antigenic than when immobilized on SA-PS. However, two monoclonals specific for the IgG2a(A2) allotypic variant, favored the adsorbed protein and one polyclonal best recognized the IgG2a(A1) variant adsorbed on Imm 2 rather than when adsorbed on PEP or immobilized on SA-PS. Both IgG1 and IgG2a, bound by apparent protein-protein interactions to an albumin monolayer, were significantly more detectable than when directly adsorbed on either Imm 2 or PEP. Using 125I-antibody or its Fab fragment to reduce steric hindrance in detection, we observed the same differences in detectability as when measured by enzyme-linked immunosorbent assay. Failure to identify a steric hindrance effect and the preference of some antibodies for adsorbed allotypic variants, support the concept of adsorption-induced conformational change (AICC). We conclude that proteins adsorbed as a monolayer on the PEP elastomer used to form the envelope of silicone breast implants are conformationally altered, but not necessarily to the same extent or the same manner as when adsorbed on polystyrene. The significantly great antigenicity of secondarily adsorbed IgG suggests that it may be present in near native conformation.
Butler
J E
JE
Department of Microbiology, University of Iowa, Iowa City 52242-1109, USA.
Navarro
P
P
Lü
E P
EP
D000327:Adsorption; D000418:Albumins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal; D016525:Antigenic Modulation; D001426:Bacterial Proteins / Q000737:chemistry; D001710:Biotin; D002417:Cattle; D004129:Dimethylpolysiloxanes / Q000737:chemistry* / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005720:Gamma Rays; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D015592:Immunoradiometric Assay / Q000379:methods; D011137:Polystyrenes / Q000737:chemistry / Q000378:metabolism / Q000528:radiation effects; D012828:Silicones / Q000737:chemistry* / Q000378:metabolism*; D019809:Streptavidin; D013499:Surface Properties
D000418:Albumins; D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D004129:Dimethylpolysiloxanes; D007074:Immunoglobulin G; D011137:Polystyrenes; D012828:Silicones; C013830:baysilon; D001710:Biotin; D019809:Streptavidin