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. 2017 Nov 3;7:14476. doi: 10.1038/s41598-017-15251-6

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Bevacizumab, Rituximab, and Trastuzuzumab Heavy Chain Variable Domain Peptide Library Screening: A peptide library of the heavy chain variable domain for Bevacizumab, Rituximab, and Trastuzumab were screened via Biacore over immobilized HSA and an HSA peptide library, as well as vice versa. Peptides were run in the pM-uM concentration range. (a) The positive results of the peptide screening. Binding kinetics between antibody variable domain peptides and HSA Peptide 40/HSA were determined by Biacore Evaluation Software analysis of SPR sensograms. (b) Multiple sequence alignment of the variable region peptides in question, performed by Clustal Omega. (c) Rituximab digest Biacore screening. Rituximab was trypsin digested and separated by HPLC reverse phase chromatography. The fraction (Fraction 58) that bound both HSA Peptide 40 and albumin was sequenced via mass spec. (d) 3D structure prediction of peptides in solution performed by PEP-FOLD 3.0, visualized in Chimera. (e) Biolayer Interfermoetry (BLITZ) assay kinetics assay of either biotinylated bevacizumab or Bev Variable Domain Peptide bound to strepdavidin probe against Abraxane nanoparticles. Pembrolizumab included as a negative control.