Journal: Communications Chemistry

Article Title: Creating unimolecular multivalent diversity in protein conjugates via the Passerini multicomponent bioconjugation with isocyanoproteins

doi: 10.1038/s42004-025-01827-1

Figure Lengend Snippet: Passerini bioconjugation for the assembly of multivalent glycoconjugates incorporating bacterial polysaccharide antigens . A Repeating unit of the pneumococcal serotype 14 (Pn14) and the meningococcal serogroup C (MenC) capsular polysaccharides (CPs). B SE-HPLC traces (TSK 5000 PW column) of the natural, fragmented and—for CPs Pn14—oxidized polysaccharides. C Schematic representation of the structures of the multivalent BSA-MenC-Pn14 glycoconjugates produced by the Passerini bioconjugation with isocyanoproteins. D Antigenicity evaluation by Dot blot assays of glycoconjugates 23, 24, 25 and 26 in comparison with isocyano-BSAs and modified CPs, using reference antibodies against BSA, S. pneumoniae and N. meningitidis . Native BSA and the natural CPs antigens were used as positive controls.

Article Snippet: Next, the membrane was incubated with the primary antibodies: anti-BSA rabbit polyclonal IgG antibody (1:1000, Invitrogen Ref A11133, Lot 2206803), anti- S. pneumoniae rabbit polyclonal IgG antibody (1:1000, Bio-Rad Ref 0300-0218, Lot 155340, reactive for serotype 14) and anti- N. meningitidis rabbit polyclonal IgG antibody (1:2000, Bio-Rad Ref 6600-5906, Lot 158003, reactive for serogroup C) in a 1% skim milk solution in TBS at 37 °C for 60 min. After washing the membrane three times for 5 min with the washing solution, it was incubated with an anti-rabbit IgG antibody conjugated to HRP (1:10 000, Santa Cruz SC2357) in TBS at 37 °C for 60 min.

Techniques: Produced, Dot Blot, Comparison, Modification