A new method for induction of specific polyclonal antibodies using immunostained proteins on nitrocellulose membrane, and HIV 1 core antigen and Escherichia coli verotoxin as examples.
Two polyclonal antibodies against verotoxin 1 and the core protein p24 of HIV 1 were raised in mice by a new immunization procedure. Both proteins were transferred to nitrocellulose, reacted with polyspecific antisera and the antigen-antibody complexes were then visualized by immunostaining. For preparation of antisera the stained protein bands were cut from the nitrocellulose sheets and implanted subcutaneously into the backs of BALB/c mice, without any adjuvant. A single booster was given 4 weeks later by implanting a second strip. All mice produced high titers of antibody directed against the antigen used for immunization. Thus, antibodies of high specificity can be elicited against protein bands in stained immunoblots.
Sandkamp
O
O
Institut für Medizinische Mikrobiologie und Immunologie der Universität Hamburg, Universitätskrankenhaus Eppendorf, F.R.G.
Karch
H
H
Laufs
R
R
D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D000917:Antibody Formation; D000918:Antibody Specificity; D001427:Bacterial Toxins / Q000276:immunology; D003101:Collodion; D004926:Escherichia coli / Q000276:immunology; D005260:Female; D015683:Gene Products, gag / Q000276:immunology; D015483:HIV Antibodies / Q000096:biosynthesis; D016655:HIV Core Protein p24; D015497:HIV-1 / Q000276:immunology; D007158:Immunologic Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D011506:Proteins / Q000276:immunology* / Q000302:isolation & purification; D022622:Shiga Toxin 1; D013194:Staining and Labeling; D014758:Viral Core Proteins / Q000276:immunology
D000907:Antibodies, Bacterial; D001427:Bacterial Toxins; D015683:Gene Products, gag; D015483:HIV Antibodies; D016655:HIV Core Protein p24; D011506:Proteins; D022622:Shiga Toxin 1; D014758:Viral Core Proteins; D003101:Collodion