In situ chemical modification of peptide microarrays: application to the study of the antibody responses to methylated antigens.
Peptide microarrays are useful tools for characterizing the humoral response against methylated antigens. They are usually prepared by printing unmodified and methylated peptides on substrates such as functionalized microscope glass slides. The preferential capture of antibodies by methylated peptides suggests the specific recognition of methylated epitopes. However, unmodified peptide epitopes can be masked due to their interaction with the substrate. The accessibility of unmodified peptides and thus the specificity of the recognition of methylated peptide epitopes can be probed using the in situ methylation procedure described here. Alternately, the in situ methylation of peptide microarrays allows probing the presence of antibodies directed toward methylated epitopes starting from easy-to-make and cost-effective unmodified peptide libraries. In situ methylation was performed using formaldehyde in the presence of sodium cyanoborohydride and nickel chloride. This chemical procedure converts lysine residues into mono- or dimethyl lysines.
Desmet
Rémi
R
UMR 8161 CNRS-Université de Lille Nord de France, Lille, France.
Diesis
Eric
E
Drobecq
Hervé
H
Rouanet
Carine
C
Chemlal
Karim
K
Debrie
Anne-Sophie
AS
Hougardy
Jean-Michel
JM
Mascart
Françoise
F
Locht
Camille
C
Melnyk
Oleg
O
D000478:Alkylation; D000588:Amines / Q000737:chemistry; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000276:immunology; D001894:Borohydrides / Q000737:chemistry; D005557:Formaldehyde / Q000737:chemistry; D005898:Glass / Q000737:chemistry; D037102:Lectins / Q000737:chemistry* / Q000276:immunology*; D008239:Lysine / Q000737:chemistry; D008745:Methylation; D051379:Mice; D008853:Microscopy; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis; D009532:Nickel / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D011327:Printing; D040081:Protein Array Analysis / Q000379:methods*
D000588:Amines; D000906:Antibodies; D001426:Bacterial Proteins; D001894:Borohydrides; D037102:Lectins; D010446:Peptide Fragments; C104578:heparin-binding hemagglutinin; D005557:Formaldehyde; C022838:nickel chloride; D009532:Nickel; C009282:sodium cyanoborohydride; D008239:Lysine