custom glass-bound peptide array consisting of 768 overlapping peptides (of 20-25 amino acid length) derived from the above-selected proteins  (INTAVIS Inc)

Journal: The Journal of Infectious Diseases

Article Title: A Protein Epitope Targeted by the Antibody Response to Kawasaki Disease

doi: 10.1093/infdis/jiaa066

Figure Lengend Snippet: Experimental strategy and characteristics of PB isolated from peripheral blood of 11 children 1–3 weeks after KD fever onset. A, Study overview. B–D, Analysis of single cells from 11 children with KD. B, Most PB were VH3, VH4, or VH1. C, Number of PB by isotype; IgA and IgG PB were most commonly identified. D, Of 1156 PB sequenced, 42 sets of oligoclonal PB were identified for antibody production and 15 somatically mutated IgA plasmablasts were also selected for production. Of 60 monoclonal antibodies prepared, 10 strongly bound to intracytoplasmic inclusion bodies in ciliated bronchial epithelial cells of children who died of KD using immunohistochemistry and 22 were found to bind weakly. Strongly binding antibodies were tested on an animal virus peptide array. Abbreviations: IgA, immunoglobulin A; IgG, immunoglobulin G; KD, Kawasaki disease; PB, plasmablasts; RT-PCR, reverse transcription and polymerase chain reaction; VH, variable heavy; VL, variable light.

Article Snippet: A custom animal virus discovery peptide array was designed (PEPperPRINT, www.pepperprint.com ) and antibodies KD4-2H4 and KD6-2B2 tested on the array.

Techniques: Isolation, Bioprocessing, Immunohistochemistry, Binding Assay, Virus, Peptide Microarray, Reverse Transcription Polymerase Chain Reaction, Reverse Transcription, Polymerase Chain Reaction

Journal: The Journal of Infectious Diseases

Article Title: A Protein Epitope Targeted by the Antibody Response to Kawasaki Disease

doi: 10.1093/infdis/jiaa066

Figure Lengend Snippet: Identifying the peptide motif recognized by Kawasaki disease (KD) monoclonal antibody KD4-2H4 using animal virus peptide array and substitution array. A, Animal virus peptide array demonstrates that KD4-2H4 recognizes multiple related peptides with averaged median foreground fluorescence intensities above a cutoff value of 200; the epitope identity number from the Immune Epitope Database (www.iedb.org) is listed for each reacting peptide. B, The motif of KD4-2H4 binding identified as statistically significant by MEME bioinformatics analysis. C, Substitution matrix array of peptide AIIPDREALYQEFDEME using KD4-2H4. Preferred amino acids for binding are those on the left of the substitution matrix while nonpreferred amino acids are those on the right.

Article Snippet: A custom animal virus discovery peptide array was designed (PEPperPRINT, www.pepperprint.com ) and antibodies KD4-2H4 and KD6-2B2 tested on the array.

Techniques: Virus, Peptide Microarray, Fluorescence, Binding Assay