High Affinity Anti-inorganic Material Antibody Generation by Integrating Graft and Evolution Technologies: POTENTIAL OF ANTIBODIES AS BIOINTERFACE MOLECULES

Takamitsu Hattori,Mitsuo Umetsu,Takeshi Nakanishi,Takanari Togashi,Nozomi Yokoo,Hiroya Abe,Satoshi Ohara,Tadafumi Adschiri,Izumi Kumagai

doi:10.1074/jbc.m109.020156

Abstract

Recent advances in molecular evolution technology enabled us to identify peptides and antibodies with affinity for inorganic materials. In the field of nanotechnology, the use of the functional peptides and antibodies should aid the construction of interface molecules designed to spontaneously link different nanomaterials; however, few material-binding antibodies, which have much higher affinity than short peptides, have been identified. Here, we generated high affinity antibodies from material-binding peptides by integrating peptide-grafting and phage-display techniques. A material-binding peptide sequence was first grafted into an appropriate loop of the complementarity determining region (CDR) of a camel-type single variable antibody fragment to create a low affinity material-binding antibody. Application of a combinatorial library approach to another CDR loop in the low affinity antibody then clearly and steadily promoted affinity for a specific material surface. Thermodynamic analysis demonstrated that the enthalpy synergistic effect from grafted and selected CDR loops drastically increased the affinity for material surface, indicating the potential of antibody scaffold for creating high affinity small interface units. We show the availability of the construction of antibodies by integrating graft and evolution technology for various inorganic materials and the potential of high affinity material-binding antibodies in biointerface applications.

Highlights

Peptides and proteins recognize the interfacial surfaces of their corresponding molecules with high affinity and selectivity because of the multiple-point interactions of hydrogen bonds and salt bridges and the surficial complementarities at the inter
We preserved the edge sequences of the complementarity determining region (CDR) when the CDR loops were replaced with the ZnO-binding peptide (ZnOBP) sequence
Single Domain Camel VHH Fragment as a Framework for Grafting Material-binding Peptide—The structural resemblance of human and mouse Fv frameworks enables the transfer of binding function to humanized antibodies [33]; these replacement techniques are currently applied to the design of new functional antibody fragments by the grafting of alien

Summary

Introduction

Peptides and proteins recognize the interfacial surfaces of their corresponding molecules with high affinity and selectivity because of the multiple-point interactions of hydrogen bonds and salt bridges and the surficial complementarities at the inter-. For the selection of VHH with high affinity for inorganic materials, chain camel antibody (VHH) to give a VHH fragment with the ϳ109 phages were mixed with 0.2 mg of material particles about same affinity as the grafted peptide and without structural 100 nm in diameter

Results

Conclusion