A Study of the Heterogeneity of Human Alpha₁-Acid Glycoprotein with Monoclonal Antibodies

Abstract

Monoclonal antibodies (MABS) were raised against human alpha 1-acid glycoprotein (AGP) and their reaction with the polymorphic forms of this plasma protein were evaluated. Spleen cells of BALB/c mice, immunized with native or desialylated human AGP, were fused with NSO mouse myeloma cells. The hybridoma products were screened with a direct ELISA test, in which the immunoplates were coated with a mixture of native and desialylated AGP. In this test, 14 anti-AGP antibody producing clones were retained. Coating the wells with either native or desialylated AGP showed that eleven clones reacted with both types of AGP ('Type I' MABS), while three MABS reacted specifically with the desialylated form ('Type II' MABS). Precoating the immunoplates with polyclonal anti-human AGP followed by incubation with native or desialylated AGP before the addition of hybridoma supernatant (indirect ELISA), confirmed the specificities observed in the direct ELISA. The molecular heterogeneity of both native AGP and desialylated AGP, based on Concanavalin A reactivity and isoelectric point, was not reflected by any specificity in the antibody reactions. Thus 'Type I' MABS reacted with all molecular forms present in native and desialylated AGP while 'Type II' MABS reacted with all molecular forms present in desialylated AGP.

Endothelium-independent vasorelaxant effect of 20(S)-protopanaxadiol on isolated rat thoracic aorta

“…Specific activation of α1 receptors induces vasoconstriction and increases blood pressure. In contrast, vascular smooth muscle relaxes after β-adrenergic receptor stimulation [28][29][30] . In this study, propranolol, an inhibitor of the β-adrenergic receptor, did not attenuate 20(S)-PPD-induced relaxation.…”

Orosomucoid Phenotyping with Monoclonal Antibodies: Polymorphic Occurrence of ORM1*Q0 in Aboriginal Taiwanese Populations

“…OR48 did not rec ognize ORM2 H19 but did label ORM2 M. The three monoclonal antibodies reacted to both native and desialylated forms of ORM (data not shown), although Fraeyman et al [11] have shown that some of the monoclonal antibodies react to both the native and the desialylated ORM, whereas others react only to the desialylated form of ORM. ORM phenotyping was performed on 658 individuals from nine subpopulations of abo riginal Taiwanese by IEF and immunoprinting using the polyclonal anti-ORM antiserum from Dako.…”