Directed evolution of polypropylene and polystyrene binding peptides
Congratulations Kristin Rübsam and Lina Weber on their recent Publication!Copyright: Bio VI
Surface functionalization of biological inert polymers, for example polypropylene and polystyrene, with material binding peptides facilitates an efficient immobilization of enzymes, bioactive peptides or antigens at ambient temperature in water. The developed Peptide Polymer evolution protocol (PePevo) is robust directed evolution protocol that enables to tailor polymer binding anchor peptides for efficient binding under application conditions. Key for a successful directed evolution campaign was to develop an epPCR protocol with a very high mutation frequency, 60 mutations per kb, to ensure sufficient diversity in the peptides LCI and Tachystatin A2. LCI and Tachystatin A2 were genetically fused to the reporter eGFP to quantify peptide binding on polypropylene and polystyrene surfaces by fluorescence analysis. PePevo was validated in two directed evolution campaigns for both peptides and polymers, LCI and polypropylene as well as Tachystatin A2 and Polystyrene. The nonionic surfactant Triton X-100, 1 mM for LCI, and the anionic surfactant LAS, 0.5 mM for Tachystatin A2, were used as selection pressure for improved peptide binders. PePevo yielded in two up to three fold improved LCI polypropylene binders, I24T, Y29H, E42K and D31V, E42G and in two up to six fold stronger Tachystatin A2 polystyrene binders R3S, L6P, V12K, S15P, C29R, R30L, F33S, Y44H and F9C, C24S, G26D, S31G, C41S, Y44Q.
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Rübsam*, K., Weber*, L., Jakob, F., Schwaneberg, U. (2017). Directed evolution of polypropylene and polystyrene binding peptides. Biotechnol. Bioeng., first published online: October 24 2017, DOI: 10.1002/bit.26481.