Isolation of a Basic Protein Antigen of Low Ragweed Pollen

Abstract

GRIFFITIIS, B. W., ~m BRUNET, R. Isolation of a basic protein antigen of low ragweed pollen, Can.J. Biochem. 49,596400 (1976).A fractionation method has been developed for the preparation of a basic protein antigen @PA-R) from ragweed pollen. This antigen possesses potent allergenic activity. It is homogeneous by gel filtration on Sephadex 6-188 and carboxymethyl-Sephadex chromatography. By the former technique the molecular weight of the BPA-R was estimated to be 28 886. Judged by its physicochemical and immunologic P roperties the BPA-R appears to be distinct from several other highly purified antigens currently isolated rssm ragweed pollen.The BPA-R and AgE (a highly purified acidic protein antigen) when tested together in Ouchterlony analysis displayed reactions of typical partial imumological identity with rabbit anti-ragweed sera which were produced by hyperimmunization utilizing complete Freund's adjuvant. In contrast rabbit antisera, produced on a shorter schedule with BPA-R (using incomplete Freund's adjuvant), failed to react by Quchterlony test with the AgE but reacted strongly with the BPA-R. Further immunological as well as chemical studies will be necessary to elucidate the antigenic relationship between these moieties.G~nm-Frnm, B. W., .ET BRUNET, R. Isolation of a basic protein antigen of low ragweed pollen. Can.J. Biochem. 49,396-488 (1976).Une mtthode de fractionnement conduisant B la preparation d'un antig6ne proteique basique @PA-R) de pollen de jacobQ a t t t mise au point. Cet arntighne est un allergtne puissant. I1 est homogtne en chromatographie sur sarboxymdthyl-Sephadex et en gel-filtration sur Sephadex G-108. Cette derni&e technique perrnet d'estimer le poids moltculaire de BPA-R i 3 28 000. Par son comportement physico-chimique et immunologique %e BPA-R semble se distinguer de plusieurs autres antigtnes hautement purifits et couramment isoles du pollen de jamb&.A 19arnalyse d'Quchterlony, le BPA-R et un antighne prottique acide hautement purifit (AgE) donnent des reactions d'identitd i m~~1 s l o g i q u e partielle typique avec les sQums anti-jacobCe de lapin, lesquels sont produits par layperimuniation B 19aide de B'adjuvant complet de Freund. Par contre, les antiskrums de lapin produits par Ie BPA-R (& l'aide de 19adjuvant incomplet de Freund) suivant un prockdt abrtgt5 sont hsapables de r h g h avec 1'AgE lorsque soumis au test d'Ouchterlony, mais ils reagissent fortement avec le BPA-R. D'autres ktudes immunologiques et chimiques devront &re faites pour klucider %a relation antbtnique entre ces protkines.Recent studies utilizing modern biochemical fractionation techniques ( 1-31 have resulted in the preparation of two highly purified acidic protein antigens AgE and AgK, both of which are potent allergens. These investigations, as well as more recent ones (Mi), indicate that ragweed pollen contains multiple allergens, but the present knowledge pertaining to immunologic and structural relationships amongst the allergens remains obscure* Studies are in progress in this laboratory to i...

Partial Purification of Antigen E from Mixed Stems and Leaves of Short Ragweed Plant

“…Other researchers reported thc purification of antigens E and K (3-5), of Ra. 3 (6), and of a basic protein from short ragweed pollen (7). The distribution of short ragweed pollen antigen E in othcr portions of this plant was investigated in this study.…”

Characteristics of Five Monoclonal Antibodies to Major Allergens of the Short Ragweed Pollen

“…King et al [20] have hypothe sized that the high allergenic activities of major aller gens, AgE and AgK, are due to their possession of de terminant groups common to other proteins in rag weed pollen. It is based on immunodiffusion experi ment results that AgE and ragweed allergen BPA-R with a molecular weight of 28 kD possess a common antigenic determinant [21]. In this study, since puri fied BPA-R was not available, it could not be deter mined if any of the components shared common anti genic determinants with AgE corresponding to this al lergen.…”

Metabolomics approaches for characterizing metabolic interactions between host and its commensal microbes

“…Therefore, the measurement of bile acids in various compartments and systemic delineation of the changes of bile acids in whole gastrointestinal tracts, tissue, and circulation are very important for the full understanding of biological functions of bile acids. To date, numerous reports have been analyzed bile acids in various biological matrices [56], such as chemical [57], thin layer chromatography [58], HPLC [59], radioimmunoassay [60], enzyme linked colorometric and radioimmunoassay [61], MS [62] and direct infusion ESI-MS [63], MS/MS [64], GC using high resolution glass capillary columns and MS [65], GC [56,66], GC-MS [67], enzymatic colorimetric method [68] and enzymatic fluorimetric method [69,70], NMR spectroscopy [71], LC coupled with ultraviolet (UV) [72] or MS [73,74], HPLC-MS/MS [75,76], HPLC-ELSD [77], and ultraperformance LC-MS detection [34,[78][79][80]. NMR methods are relatively insensitive and only applicable in specific areas where the concentration of bile acids is very high, such as the analysis of bile composition [81].…”