Antibody response in immunized rabbits measured with bacterial immunoglobulin-binding proteins

Forskningsoutput: TidskriftsbidragArtikel i vetenskaplig tidskrift

Standard

Antibody response in immunized rabbits measured with bacterial immunoglobulin-binding proteins. / Elbashir, M I; Nilson, B H; Akesson, P; Björck, L; Akerström, B.

I: Journal of Immunological Methods, Vol. 135, Nr. 1-2, 31.12.1990, s. 171-9.

Forskningsoutput: TidskriftsbidragArtikel i vetenskaplig tidskrift

Harvard

APA

CBE

MLA

Vancouver

Author

RIS

TY - JOUR

T1 - Antibody response in immunized rabbits measured with bacterial immunoglobulin-binding proteins

AU - Elbashir, M I

AU - Nilson, B H

AU - Akesson, P

AU - Björck, L

AU - Akerström, B

PY - 1990/12/31

Y1 - 1990/12/31

N2 - Protein G, an immunoglobulin (Ig)-binding protein isolated from group C or G streptococci, binds to the Fc portion of IgG. Protein L, from the anaerobic bacterium Peptostreptococcus magnus, specifically binds light chains of Ig. In this study, protein G and L were used to measure the production of antibodies in immunized rabbits. Two rabbits were immunized with a mixture of human urinary proteins from a patient with tubular proteinuria, and blood samples were collected regularly from the animals for 6 weeks after the immunization. The antibody levels of the blood samples against six of the proteins in the antigen mixture were then measured by ELISA. Microtiter plates were coated with each of the antigens, incubated with the rabbit serum samples, and the specific antibodies of the IgG class measured by incubation with biotinylated protein G, and antibodies of all Ig classes with biotinylated protein L. Alternatively, Western blotting was employed, where the antibodies which bound to each antigen after separation by SDS-PAGE and transfer to nitrocellulose membranes, were detected by protein G or L. The results showed that antibody production against five of the antigens, albumin, alpha 1 gamma-acid glycoprotein, alpha 1 gamma-microglobulin, Ig light chains, and retinol-binding protein, showed a similar pattern, although the magnitude of the initial IgM response differed somewhat. After 6 weeks, the levels of the protein G-binding antibodies had reached a plateau, while those of protein L-binding antibodies were still increasing. The response to the sixth antigen, beta 2 microglobulin, was considerably different. A dramatic increase of anti-beta 2 gamma-microglobulin antibodies was seen during the 4th week after immunization when protein L was used.

AB - Protein G, an immunoglobulin (Ig)-binding protein isolated from group C or G streptococci, binds to the Fc portion of IgG. Protein L, from the anaerobic bacterium Peptostreptococcus magnus, specifically binds light chains of Ig. In this study, protein G and L were used to measure the production of antibodies in immunized rabbits. Two rabbits were immunized with a mixture of human urinary proteins from a patient with tubular proteinuria, and blood samples were collected regularly from the animals for 6 weeks after the immunization. The antibody levels of the blood samples against six of the proteins in the antigen mixture were then measured by ELISA. Microtiter plates were coated with each of the antigens, incubated with the rabbit serum samples, and the specific antibodies of the IgG class measured by incubation with biotinylated protein G, and antibodies of all Ig classes with biotinylated protein L. Alternatively, Western blotting was employed, where the antibodies which bound to each antigen after separation by SDS-PAGE and transfer to nitrocellulose membranes, were detected by protein G or L. The results showed that antibody production against five of the antigens, albumin, alpha 1 gamma-acid glycoprotein, alpha 1 gamma-microglobulin, Ig light chains, and retinol-binding protein, showed a similar pattern, although the magnitude of the initial IgM response differed somewhat. After 6 weeks, the levels of the protein G-binding antibodies had reached a plateau, while those of protein L-binding antibodies were still increasing. The response to the sixth antigen, beta 2 microglobulin, was considerably different. A dramatic increase of anti-beta 2 gamma-microglobulin antibodies was seen during the 4th week after immunization when protein L was used.

KW - Animals

KW - Antibody Formation

KW - Bacterial Proteins

KW - Biotin

KW - Blotting, Western

KW - Electrophoresis, Polyacrylamide Gel

KW - Enzyme-Linked Immunosorbent Assay

KW - Immunization

KW - Immunoglobulin Fc Fragments

KW - Immunologic Techniques

KW - Proteins

KW - Proteinuria

KW - Rabbits

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1016/0022-1759(90)90270-6

DO - 10.1016/0022-1759(90)90270-6

M3 - Article

C2 - 2273256

VL - 135

SP - 171

EP - 179

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

SN - 1872-7905

IS - 1-2

ER -