{"id":9756,"date":"2022-09-02T06:00:07","date_gmt":"2022-09-01T21:00:07","guid":{"rendered":"https:\/\/www.emukk.com\/WP\/?p=9756"},"modified":"2022-09-02T06:54:00","modified_gmt":"2022-09-01T21:54:00","slug":"a-novel-monoclonal-igg1-antibody-specific-for-%ce%b1-gal-epitope","status":"publish","type":"post","link":"https:\/\/www.emukk.com\/WP\/en\/a-novel-monoclonal-igg1-antibody-specific-for-%ce%b1-gal-epitope\/","title":{"rendered":"A novel monoclonal IgG1 antibody specific for \u03b1-Gal epitope"},"content":{"rendered":"<p>A group from Technical University of Munich, School of Medicine, Neuherberg, Germany, etc. has reported about a novel monoclonal IgG1 antibody specific for \u03b1-Gal epitope.<br \/>\n<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC9391071\/\">https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC9391071\/<\/a><\/p>\n<p>The alpha-Gal (\u03b1-Gal) epitope is a carbohydrate immunogen in humans that has relevance in allergy and xenotransplantation. Interestingly, antibodies of different isotypes against the \u03b1-Gal epitope are quite abundant in humans with IgG levels estimated to range between 1% to 0.1% of total plasma IgG with high variability between subjects and lowest abundance in individuals carrying the blood type B antigen. This observation is likely due to the structural similarity between the \u03b1-Gal epitope and blood type B antigen, which contains an additional fucose molecule on the second last galactose molecule. These human anti-\u03b1-Gal antibodies pose a challenge for xenotransplantation, in particular for pig organ transplantation, which was overcome to some extend with developing GGTA1 knockout (KO) pigs.<\/p>\n<p>In this paper, the development of a novel IgG1 antibody called 27H8 was reported, which is highly specific for both synthetic and naturally occurring \u03b1-Gal epitopes. In order to generate a monoclonal antibody specific for the \u03b1-Gal epitope determining structure Gal-\u03b11,3-Gal that is equally able to bind to the naturally occurring \u03b1-Gal epitope Gal-\u03b11,3-Gal-\u03b21,4-GlcNAc, \u03b1-galactosyltransferase knockout mice (Ggta1 KO) were immunized with Gal-\u03b11,3-Gal-\u03b21,4-GlcNAc coupled to ovalbumin as carrier protein (\u03b1-Gal-OVA).<\/p>\n<p>To verify the specificity, the 27H8 monoclonal antibody was compared to Bandereia simplifolica isolectin B4 (BSI-B4: GSL-I B4) and to the monoclonal IgM antibody M86, which are both widely used to detect the \u03b1-Gal epitope. BSI-B4 is specific for terminal \u03b1-galactose oligosaccharides and therefore recognizes also the blood group B antigen, which differs from the \u03b1-Gal epitope only in the addition of one fucose residue and is thus structurally very similar. To assess whether 27H8 also binds to the blood group B antigen we blotted lysates of whole blood from a type B donor on a membrane and applied the antibodies 27H8 and M86 or biotinylated BSI-B4 for detection. While BSI-B4 bound to the blood type B specimen as expected, neither 27H8 or M86 did. Next, it was investigated whether 27H8 also binds to natural \u03b1-Gal epitopes. As pig kidney is naturally rich in \u03b1-Gal and reactions in \u03b1-Gal allergic patients are severe after ingestion, it was also tested if 27H8 recognizes \u03b1-Gal in pig kidney lysates in a dot blot assay. 27H8 binding to wildtype (WT) pig kidney lysate was observed with strong staining intensity.\u3000However, there was no signal on a sample digested with \u03b1-Galactosidase (Dig. WT).<\/p>\n<p><a href=\"https:\/\/www.emukk.com\/WP\/wp-content\/uploads\/2022\/09\/27H8-specificity-to-a-Gal.png\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone size-medium wp-image-9753\" src=\"https:\/\/www.emukk.com\/WP\/wp-content\/uploads\/2022\/09\/27H8-specificity-to-a-Gal-300x152.png\" alt=\"\" width=\"300\" height=\"152\" srcset=\"https:\/\/www.emukk.com\/WP\/wp-content\/uploads\/2022\/09\/27H8-specificity-to-a-Gal-300x152.png 300w, https:\/\/www.emukk.com\/WP\/wp-content\/uploads\/2022\/09\/27H8-specificity-to-a-Gal-600x305.png 600w, https:\/\/www.emukk.com\/WP\/wp-content\/uploads\/2022\/09\/27H8-specificity-to-a-Gal-1024x520.png 1024w, https:\/\/www.emukk.com\/WP\/wp-content\/uploads\/2022\/09\/27H8-specificity-to-a-Gal-768x390.png 768w, https:\/\/www.emukk.com\/WP\/wp-content\/uploads\/2022\/09\/27H8-specificity-to-a-Gal-920x467.png 920w, https:\/\/www.emukk.com\/WP\/wp-content\/uploads\/2022\/09\/27H8-specificity-to-a-Gal-230x117.png 230w, https:\/\/www.emukk.com\/WP\/wp-content\/uploads\/2022\/09\/27H8-specificity-to-a-Gal-350x178.png 350w, https:\/\/www.emukk.com\/WP\/wp-content\/uploads\/2022\/09\/27H8-specificity-to-a-Gal-480x244.png 480w, https:\/\/www.emukk.com\/WP\/wp-content\/uploads\/2022\/09\/27H8-specificity-to-a-Gal.png 1059w\" sizes=\"auto, (max-width: 300px) 100vw, 300px\" \/><\/a>\u3000refer to the original paper for detailed explanation<\/p>\n","protected":false},"excerpt":{"rendered":"<p>A group from Technical University of Munich, School of Medicine, Neuherberg, Germany, etc. has reported about <\/p><\/div>\n<div class=\"blog-btn\"><a href=\"https:\/\/www.emukk.com\/WP\/en\/a-novel-monoclonal-igg1-antibody-specific-for-%ce%b1-gal-epitope\/\" class=\"home-blog-btn\">\u7d9a\u304d\u3092\u8aad\u3080<\/a><\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[7],"tags":[],"class_list":["post-9756","post","type-post","status-publish","format-standard","hentry","category-technology-en"],"aioseo_notices":[],"_links":{"self":[{"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/posts\/9756","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/comments?post=9756"}],"version-history":[{"count":3,"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/posts\/9756\/revisions"}],"predecessor-version":[{"id":9791,"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/posts\/9756\/revisions\/9791"}],"wp:attachment":[{"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/media?parent=9756"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/categories?post=9756"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/tags?post=9756"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}