Influence of target cell preparation on binding of monoclonal islet cell reactive antibodies (mc-ICRA) in cellular enzyme-linked immunosorbent assay (CELISA)

Schlosser, Michael; Witt, Sabine; Ziegler, B; Ziegler, M.

doi:10.1016/0022-1759(91)90131-x

Cited by 19 publications

(6 citation statements)

References 23 publications

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“…The time course of the serum IgG contents of rats was determined by sandwich enzyme‐linked immunosorbent assay (ELISA)14 with specific solid‐phase‐coated antisera (rabbit antirat IgG, Jackson ImmunoResearch Laboratories, Inc.), rat immunoglobulin reference sera as a standard (ICN Biomedicals GmbH, Eschwege, Germany), and peroxidase conjugated rabbit antirat IgG (Fc‐γ‐chain‐specific) as described for polymer antibodies.…”

Section: Methodsmentioning

confidence: 99%

Immunogenicity of polymeric implants: Long‐term antibody response against polyester (Dacron) following the implantation of vascular prostheses into LEW.1A rats

Schlosser

Wilhelm

Gunnarsson³

et al. 2002

J. Biomed. Mater. Res.

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Implanted biomaterials trigger acute and chronic inflammatory responses directly correlated to the central role of phagocytic cells at the host-implant interface. This study was designed to evaluate specific humoral immune responses following repeated intraperitoneal implantations of collagen-impregnated polyester (Dacron) prosthetic segments into LEWIS rats. Serum antibody detection was performed by enzyme immunoassay with the prosthetic segments as a target. Cutoff values for antibody positivity were greater than or equal to the 99th percentile for control rats. Polymer immunoglobiulin G (IgG) antibodies were significantly increased (p < 0.05) by repeated implantation and were subsequently followed until experimental day 293. Antibody formation was significantly enhanced through the application of complete Freund's adjuvant in combination with the first implantation. All rats within this group were antibody-positive on day 53, but only 6 of 10 animals that received the prosthesis without the adjuvant were. After preincubation of sera with bovine collagen type I (solid phase adsorbed or in solution), polymer antibody binding was discovered not to be diminished, indicating that the IgG antibodies detected were not directed against the prosthesis impregnation. Furthermore, a significant correlation was obtained between polymer antibody binding to collagen-impregnated and nonimpregnated prostheses (r(s) = 0.797, p < 0.001). There was no substantiated correlation between antibody binding to polyester and to an irrelevant polymer (Tecoflex EG 80). We conclude that specific polymer antibodies may indeed provide an additional parameter for biocompatibility testing as well as a possible serological marker of an inflammatory response to implants.

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Section: Methodsmentioning

confidence: 99%

Immunogenicity of polymeric implants: Long‐term antibody response against polyester (Dacron) following the implantation of vascular prostheses into LEW.1A rats

Schlosser

Wilhelm

Gunnarsson³

et al. 2002

J. Biomed. Mater. Res.

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“…The optical density (OD.) at 490 nm was measured using an autoreader (BIO-TEK Instruments, Winooski, VT, USA) (Schlosser et al, 1991). The inter-assay precision of the ELISA expressed as the coefficient of variation (cv) over three assays with different GAD preparations ranged from 9-17% for DP BBIOK rat sera; the intra-assay cv for duplicate determinations was 7.8%.…”

Section: Animalsmentioning

confidence: 99%

Autoantibodies to glutamate decarboxylase detected in diabetes-prone BB/OK rats do not distinguish onset of diabetes

Ziegler

Schlosser²,

Hamann³

et al. 2009

Exp Clin Endocrinol Diabetes

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The diabetes syndrome of the BB rat resembles human Type 1 (insulin-dependent) diabetes including the prevalence of autoantibodies to the 64 kDa Beta-cell autoantigen, which has been identified as glutamate decarboxylase. This study aimed at detecting the prevalence and level of glutamate decarboxylase autoantibodies in 120-day-old diabetic and non-diabetic diabetes-prone BB/OK rats compared to those of sex- and age-matched diabetes-resistant LEW.1A rats. The antibodies were detected using semipurified glutamate decarboxylase from rat brain in two immunoassays, a direct and a sandwich enzyme-linked immunosorbent assay. For the last assay autoantibody-containing immunoglobulins of a serum from a patient with the stiff-man syndrome were used to bind specifically the enzyme as autoantigen in plastic wells. The antibody levels measured as optical density at 490 nm (x +/- SD)/prevalence of the diabetic group (120 +/- 29 days of age) of BB/OK rats 0.57 +/- 0.29 (n = 51)/88% as well as those of the nondiabetic group (121 +/- 26 days of age) with 0.51 +/- 0.29 (n = 32)/97% was significantly increased (p < 0.01) compared to those of the diabetes-resistant control group 0.15 +/- 0.06 (n = 29)/0%. Furthermore in a 209 +/- 27-day-old group (n = 21) of non-diabetic but diabetes-prone BB/OK rats the autoantibody levels of 1.21 +/- 0.39 vs 0.51 +/- 0.26 were further significantly enhanced (p < 0.01). These results were confirmed by a sandwich assay.(ABSTRACT TRUNCATED AT 250 WORDS)

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“…Cell-based ELISA (cb-ELISA) using whole cells as target antigens in screening assays have been described previously, (9) including glutaraldehyde-or formalin-fixed adherent cells (10) and non-adherent cells, (11) as well as viable live adherent cells. (12) Fixed plant cells have also been used in a cb-ELISA. (13,14) However, our modified cb-ELISA methodology substantially extends its use by being able to detect hybridomas secreting MAb that react to cell surface and/or intracellular antigens.…”

Section: Amentioning

confidence: 99%

Development of a Quantitative Cell-Based Intracellular ELISA for the Screening of B Cell Hybridoma Supernatants: A Novel Rapid Assay to Detect Positive Clones

Renukaradhya

Sriram

Poláková

et al. 2004

Hybridoma and Hybridomics

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The primary screening of hybridoma clones secreting monoclonal antibodies (MAbs) requires the testing of a large number of hybridoma culture supernatants within a short time and is very labor-intensive. In addition, the type of antigen and its location in the cell have to be considered when selecting the appropriate screening procedure, but relatively few reagents are available for analyzing these molecules. We have developed an intracellular and cell surface ELISA technique for screening hybridoma supernatants that hastens the screening procedure of a large number of clones in a short period of time, as the supernatants of fused cells grown in 96-well plates are used directly in the assay. This novel screening technique is rapid, sensitive, specific, and applicable to MAbs specific for a wide variety of intracellular and/or cell surface proteins.

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Influence of target cell preparation on binding of monoclonal islet cell reactive antibodies (mc-ICRA) in cellular enzyme-linked immunosorbent assay (CELISA)

Cited by 19 publications

References 23 publications

Immunogenicity of polymeric implants: Long‐term antibody response against polyester (Dacron) following the implantation of vascular prostheses into LEW.1A rats

Immunogenicity of polymeric implants: Long‐term antibody response against polyester (Dacron) following the implantation of vascular prostheses into LEW.1A rats

Autoantibodies to glutamate decarboxylase detected in diabetes-prone BB/OK rats do not distinguish onset of diabetes

Development of a Quantitative Cell-Based Intracellular ELISA for the Screening of B Cell Hybridoma Supernatants: A Novel Rapid Assay to Detect Positive Clones

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