Selective Capture of Anti‐N‐glucosylated NTHi Adhesin Peptide Antibodies by a Multivalent Dextran Conjugate. (6th December 2021)

• Record Type: Journal Article
• Authors: Mazzoleni, Antonio; Real‐Fernandez, Feliciana; Nuti, Francesca; Lanzillo, Roberta; Brescia Morra, Vincenzo; Dambruoso, Paolo; Bertoldo, Monica; Rovero, Paolo; Mallet, Jean‐Maurice; Papini, Anna Maria
• Is Part Of: Chembiochem. Volume 23:Number 3(2022)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2021-12-06
• Subjects: antibody caption -- dextran conjugates -- ELISA -- multivalence -- peptides; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.202100515
• Languages: English
• ISSNs: 1439-4227

Abstract

Abstract: Tentacle‐like polymers decorated with several copies of peptide antigens can be interesting tools for increasing the ability to capture circulating antibodies in patient sera, using cooperative effects for stronger avidity. We previously showed that antibodies from multiple sclerosis (MS) patient sera preferentially recognize hyperglucosylated adhesin protein HMW1ct of non‐typeable Haemophilus influenzae ( NTHi ). We selected the C‐terminal HMW1ct(1347–1354) minimal epitope and prepared the diglucosylated analogue Ac‐KAN(Glc)VTLN(Glc)TTG‐K(N3 )‐NH2 to graft a 40 kDa dextran scaffold modified with glycidyl‐propargyl moieties to perform a copper catalyzed alkyne‐azide coupling reaction (CuAAC). Quantitative NMR measurements allowed the characterization of the peptide loading (19.5 %) on the multivalent dextran conjugate. This novel polymeric structure displayed optimal capturing properties of both IgG and, more interestingly, IgM antibodies in MS sera. Specific antibodies from a representative MS serum, were successfully depleted using a Sepharose resin bearing the new glucosylated multivalent conjugate, as confirmed by ELISA. These results may offer a promising proof‐of‐concept for the selective purification of high affinity autoantibodies from sera of autoimmune patients, in general, and of specific high affinity antibodies against a minimally glcosylated epitope Asn(Glc) from sera of multiple sclerosis (MS) patients, in particular. Abstract : Dextran‐based tentacles were decorated with peptide antigens as a way to increase the ability to capture circulating IgM antibodies in multiple sclerosis patient sera as disease biomarkers. A novel peptide‐dextran conjugate bearing multiple copies of a selected epitope of the non‐typeable Haemophilus Influenzae adhesin protein was synthesis and characterized. Immunological experiments provide the proof‐of concept for the identification and capture of specific IgM antibodies in patient sera.