Discovery of a polystyrene binding peptide isolated from phage display library and its application in peptide immobilization

Abstract

Phage peptide display is a powerful technique for discovery of various target-specific ligands. However, target-unrelated peptides can often be obtained and cause ambiguous results. Peptide PB-TUP has been isolated repeatedly in our laboratory on different targets and we conducted a research on PB-TUP phage to investigate their binding properties and rate of propagation. ELISA and phage recovery assay demonstrated that PB-TUP phage had a significant superior affinity to polystyrene solid surface compared with control phage clones. In this study, some incidental bindings are excluded like blocking agents and non-specific binding of secondary antibodies. Propagation rate assays of the selected phage clones showed that the growth rate of PB-TUP phage was not superior to the control phages. Furthermore, the binding of PB-TUB to polystyrene was concentration dependent and varied with solution pH. Molecular modeling revealed that stable structures of α-helix and β-turn may contribute to the binding of PB-TUP to polystyrene plate. The PB-TUP sequence was fused to the N-terminus of peptide P2 and the fusion peptide significantly increased the binding affinity to polystyrene. The fusion peptide also enhanced the cell adhesion ability of peptide P2 with human umbilical vein endothelial cell (HUVEC). The addition of the polystyrene binding peptide provided a convenient method for peptide immobilization.

Matter‐tag: A universal immobilization platform for enzymes on polymers, metals, and silicon‐based materials

“…Besides natural substrates, the adhesion of CBMs to synthetic polymers, for example, polyethylene terephthalate (PET), was reported (Ribitsch et al, 2013;Zhang, Wang, Chen, & Wu, 2013). Further, adhesion-promoting peptides are reported for numerous materials, for instance, for gold (e.g., Midas-2: TGTSVLIATPYV; Kim et al, 2010;GBP1: MHGKTQATSGTIQS; Brown, 1997;Tamerler, Oren, Duman, Venkatasubramanian, & Sarikaya, 2006;AuBP1: WAGAKRLVLRRE;Hnilova et al, 2008;AuBP2: WALRRSIRRQSY;Hnilova et al, 2008; Cys-BP: CKPKEL-PEKLPELKPK(Ahx-Ahx-Biotin)C-NH2; Zernia et al, 2016), stainless steel (e.g., MS-S1: ATIHDAFYSAPE; Zuo, Örnek, & Wood, 2005; MS-S2: NLNPNTASAMHV; Zuo et al, 2005;MTWDPSLASPRS;Vreuls et al, 2010), silicon (e.g., TBP-1: RKLPDAPGMHTW; Sano, Sasaki, & Shiba, 2005; P2:LLADTTHHRPWT; Estephan et al, 2011;Ramakrishnan, Martin, Cloitre, Firlej, & Gergely, 2014;P3: SPGLSLVSHMQT;Estephan et al, 2011;Ramakrishnan et al, 2014;PC4: SLVSHMQT;Ramakrishnan et al, 2015), polystyrene (PS; e.g., PB-TUB: VHWDFRQWWQPS ;Qiang et al, 2017;PS-19: RAFIASRRIKRP;Kumada et al, 2006;c02: YLTMPTP;Serizawa, Techawanitchai, & Matsuno, 2007; Tachystatin A2 (TA2): YSRCQLQGFNCVVRSYGLPTIPCCRGLTCRSYFPGSTYGRCQRY; Rübsam, Weber, Jakob, & Schwaneberg, 2018), and polypropylene (PP; e.g., TSDIKSRSPHHR, HTQNMRMYEPWF; Cunningham, Lowe, O'brien, Wang, & Wilkins, 2011; Cecropin A (CecA): KWKLFKKIEKVGQNIRD-GIIKAGPAVAVVGQATQIAK; Rübsam, Stomps, Böker, Jakob, & Schwaneberg, 2017; liquid chromatography peak I (LCI): AIKLVQSPNGN-FAASFVLDGTKWIFKSKYYDSSKGYWVGIYEVWDRK; Rübsam et al, 2017).…”

PSBinder: A Web Service for Predicting Polystyrene Surface-Binding Peptides

“…In addition, after the completion of our predictor, a paper published very recently reported a PSBP with the sequence of VHWDFRQWWQPS [ 40 ]. As the paper reported, this sequence does not have typical PS-binding motifs.…”

Effects of Different Broth Enrichment upon Phage Magnetoelastic Biosensor for Fast Detecting Low <i>Salmonella</i> Counts on Problematic Produce

“…Therefore, it is curious to know whether the affinity actions of the phages to Salmonella was still performed normally in the same broths, but different produce types. In the report of Qiang et al, in 2017 [25], proteins and components in different blocking buffers would inhibit the affinity of phage probes to its target and promote the non-specific binding in ELISA assays. To answer this question, Salmonella capture rates of the phage biosensors were studies.…”