4.4 Article

Selective Capture of Anti-N-glucosylated NTHi Adhesin Peptide Antibodies by a Multivalent Dextran Conjugate

期刊

CHEMBIOCHEM
卷 23, 期 3, 页码 -

出版社

WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.202100515

关键词

antibody caption; dextran conjugates; ELISA; multivalence; peptides

资金

  1. French-Italian University [C3-24]
  2. Regione Toscana, POR CReO FESR 2014-2020 (project MARK) [85438]
  3. Fondazione Ente Cassa di Risparmio Firenze [2014.0306]
  4. Italian Multiple Sclerosis Foundation (FISM) [2017/R/5]
  5. Universita degli Studi di Firenze within the CRUI-CARE Agreement

向作者/读者索取更多资源

The use of specific polymeric structures can successfully capture specific antibodies in the sera of patients with multiple sclerosis, offering a promising proof-of-concept for selectively purifying high affinity autoantibodies from the sera of autoimmune patients.
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(N-3)-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.

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