Data from: Identification and mapping of linear antibody epitopes in human serum albumin using high-density peptide arrays

We have recently developed a high-density photolithographic, peptide array
technology with a theoretical upper limit of 2 million different peptides
per array of 2 cm2. Here, we have used this to perform complete and
exhaustive analyses of linear B cell epitopes of a medium sized protein
target using human serum albumin (HSA) as an example. All possible
overlapping 15-mers from HSA were synthesized and probed with a
commercially available polyclonal rabbit anti-HSA antibody preparation. To
allow for identification of even the weakest epitopes and at the same time
perform a detailed characterization of key residues involved in antibody
binding, the array also included complete single substitution scans (i.e.
including each of the 20 common amino acids) at each position of each
15-mer peptide. As specificity controls, all possible 15-mer peptides from
bovine serum albumin (BSA) and from rabbit serum albumin (RSA) were
included as well. The resulting layout contained more than 200.000 peptide
fields and could be synthesized in a single array on a microscope slide.
More than 20 linear epitope candidates were identified and characterized
at high resolution i.e. identifying which amino acids in which positions
were needed, or not needed, for antibody interaction. As expected,
moderate cross-reaction with some peptides in BSA was identified whereas
no cross-reaction was observed with peptides from RSA. We conclude that
high-density peptide microarrays are a very powerful methodology to
identify and characterize linear antibody epitopes, and should advance
detailed description of individual specificities at the single antibody
level as well as serologic analysis at the proteome-wide level.