Selective Protein Capture by Epitope Imprinting

Summary

Principal Investigator: KENNETH SHEA
Affiliation: University of California
Country: USA
Abstract: DESCRIPTION (provided by applicant): Biological macromolecules such as proteins and DNA are essential for every life form on earth. Studies of these molecules are dependent on our ability to selectively capture them from complex biological mixtures. Antibodies have been the most widely used for selective protein and peptide capture with applications for industrial protein purification, basic biomedical research and in clinical diagnostics. Antibodies however exhibit characteristics that limit their applications. This proposal is involved with developing robust synthetic polymers for selective capture of peptides and proteins. These substances have important applications for separations, for use in biosensors, neutralization of toxins and for the development of biomedical diagnostics. The non- biological approach of molecular imprinting is used to create specific recognition sites in robust network polymers. Protein and peptide recognition is achieved by identifying an exposed domain (epitope) of the target protein, a unique nine amino acid sequence. The peptide epitope is used as the imprint molecule. We are developing two general methods for preparing imprinted polymers for protein and peptide capture, imprinted polymer films and nanosize imprinted polymer particles. Imprinted films are prepared by covalently attaching the peptide epitope to a glass or silicon surface. Monomers are then polymerized on these surfaces to produce a molecularly imprinted polymer film (MIP). Following separation from the functionalized surface, the polymer film is evaluated for its ability to capture the target protein from protein mixtures. Two methods for the preparation of MIP nanoparticles are being developed, precipitation polymerization and suspension polymerization. In these systems, epitopes are introduced with monomers in the polymerization reaction. Following isolation and dialysis, the nanoparticles are evaluated for protein and peptide affinity and specificity. In both polymer formats, films and nanoparticles, the capture is achieved under native conditions. Molecular imprinting is one of the few general, non-biological methods for creating molecular receptors. The choice of short epitopes focuses on developing capture agents for the primary structure of the peptide rather than the more complex secondary and tertiary structure of a target protein and is similar to the use of peptide fragments to generate epitope selective antibodies and synthetic materials. In addition, the capture conditions were designed to be compatible with the native protein structure. It utilizes the sequences of exposed epitopes based on known or predicted protein structure. This method requires only the peptide sequence of a small portion of the target protein for the template molecule;it does not use or need whole protein. As such, this approach provides opportunities for the capture of target proteins based only on genomic information. PUBLIC HEALTH RELEVANCE: Antibodies are important reagents that are used in biomedical research, in diagnosis of diseases, and in treatment of such diseases as infections and cancer. Antibodies are produced by cell lines or clones obtained from animals that have been immunized with the substance that is the target of study. We propose to develop a method for producing robust, inexpensive, non-biological polymer antibodies that can be used as substitutes for native antibodies.
Funding Period: 2009-01-01 - 2012-12-31
more information: NIH RePORT

Top Publications

  1. pmc Epitope discovery for a synthetic polymer nanoparticle: a new strategy for developing a peptide tag
    Keiichi Yoshimatsu
    Department of Chemistry, University of California, Irvine, California 92697, United States
    J Am Chem Soc 136:1194-7. 2014
  2. doi Polymer nanoparticle-protein interface. Evaluation of the contribution of positively charged functional groups to protein affinity
    Yusuke Yonamine
    Department of Chemistry, University of California Irvine, Irvine, California 92697, USA
    ACS Appl Mater Interfaces 5:374-9. 2013
  3. pmc Engineered synthetic polymer nanoparticles as IgG affinity ligands
    Shih Hui Lee
    Department of Chemistry, University of California, Irvine, Irvine, California 92697, United States
    J Am Chem Soc 134:15765-72. 2012
  4. pmc ELISA-mimic screen for synthetic polymer nanoparticles with high affinity to target proteins
    Yusuke Yonamine
    Department of Chemistry, University of California Irvine, Irvine, California 92697, USA
    Biomacromolecules 13:2952-7. 2012
  5. pmc Temperature-Responsive "Catch and Release" of Proteins by using Multifunctional Polymer-Based Nanoparticles
    Keiichi Yoshimatsu
    Department of Chemistry, University of California, Irvine, Irvine, CA 92697 USA
    Angew Chem Int Ed Engl 51:2405-8. 2012
  6. pmc Synthetic polymer nanoparticle-polysaccharide interactions: a systematic study
    Zhiyang Zeng
    Department of Chemistry, University of California, Irvine, Irvine, California 92697, USA
    J Am Chem Soc 134:2681-90. 2012
  7. pmc The rational design of a synthetic polymer nanoparticle that neutralizes a toxic peptide in vivo
    Yu Hoshino
    Department of Chemical Engineering, Kyushu University, 744 Motooka, Fukuoka 819 0395, Japan
    Proc Natl Acad Sci U S A 109:33-8. 2012
  8. pmc Affinity purification of multifunctional polymer nanoparticles
    Yu Hoshino
    Department of Chemistry, University of California, Irvine, California 92697, USA
    J Am Chem Soc 132:13648-50. 2010
  9. pmc Recognition, neutralization, and clearance of target peptides in the bloodstream of living mice by molecularly imprinted polymer nanoparticles: a plastic antibody
    Yu Hoshino
    Department of Chemistry, University of California, Irvine, California 92697, USA
    J Am Chem Soc 132:6644-5. 2010
  10. pmc Synthetic polymer nanoparticles with antibody-like affinity for a hydrophilic peptide
    Zhiyang Zeng
    Department of Chemistry, University of California, Irvine, California 92697, USA
    ACS Nano 4:199-204. 2010

Scientific Experts

  • Yu Hoshino
  • Kenneth J Shea
  • Yusuke Yonamine
  • Keiichi Yoshimatsu
  • Zhiyang Zeng
  • Shih Hui Lee
  • John M Beierle
  • Philip Tagari
  • Tomohiko Yamazaki
  • Les P Miranda
  • Linda F Epstein
  • Paul E Rose
  • Yoshio Okahata
  • Piere Baldi
  • Mingdi Yan
  • Ruey An Doong
  • Anuradha Tyagi
  • Benjamin K Lesel
  • Monica McCallum
  • Arlo Randall
  • Jiten Patel
  • Hoseong Yoo
  • Andy Rodriguez

Detail Information

Publications11

  1. pmc Epitope discovery for a synthetic polymer nanoparticle: a new strategy for developing a peptide tag
    Keiichi Yoshimatsu
    Department of Chemistry, University of California, Irvine, California 92697, United States
    J Am Chem Soc 136:1194-7. 2014
    ..This strategy makes available inexpensive, abiotic synthetic polymers as affinity agents for peptide tags and provides alternatives for important applications where more costly affinity agents are used. ..
  2. doi Polymer nanoparticle-protein interface. Evaluation of the contribution of positively charged functional groups to protein affinity
    Yusuke Yonamine
    Department of Chemistry, University of California Irvine, Irvine, California 92697, USA
    ACS Appl Mater Interfaces 5:374-9. 2013
    ..Circular dichroism (CD) revealed that the protein was not denatured in the process of binding or release...
  3. pmc Engineered synthetic polymer nanoparticles as IgG affinity ligands
    Shih Hui Lee
    Department of Chemistry, University of California, Irvine, Irvine, California 92697, United States
    J Am Chem Soc 134:15765-72. 2012
    ..5-8.5 nM) to the Fc domain at pH 5.5 is comparable to protein A at pH 7. These results establish that engineered synthetic polymer NPs can be formulated with an intrinsic affinity to a specific domain of a large biomacromolecule...
  4. pmc ELISA-mimic screen for synthetic polymer nanoparticles with high affinity to target proteins
    Yusuke Yonamine
    Department of Chemistry, University of California Irvine, Irvine, California 92697, USA
    Biomacromolecules 13:2952-7. 2012
    ....
  5. pmc Temperature-Responsive "Catch and Release" of Proteins by using Multifunctional Polymer-Based Nanoparticles
    Keiichi Yoshimatsu
    Department of Chemistry, University of California, Irvine, Irvine, CA 92697 USA
    Angew Chem Int Ed Engl 51:2405-8. 2012
    ..The process is reversible and does not denature the proteins. An optimized combination of functional monomers imparts binding selectivity toward a target protein over other proteins...
  6. pmc Synthetic polymer nanoparticle-polysaccharide interactions: a systematic study
    Zhiyang Zeng
    Department of Chemistry, University of California, Irvine, Irvine, California 92697, USA
    J Am Chem Soc 134:2681-90. 2012
    ..Select synthetic polymer nanoparticles have also been shown to inhibit protein-heparin interactions and thus offer the potential for therapeutic applications...
  7. pmc The rational design of a synthetic polymer nanoparticle that neutralizes a toxic peptide in vivo
    Yu Hoshino
    Department of Chemical Engineering, Kyushu University, 744 Motooka, Fukuoka 819 0395, Japan
    Proc Natl Acad Sci U S A 109:33-8. 2012
    ..These results provide a platform to design plastic antidotes and reveal the potential and possible limitations of using synthetic polymer nanoparticles as plastic antidotes...
  8. pmc Affinity purification of multifunctional polymer nanoparticles
    Yu Hoshino
    Department of Chemistry, University of California, Irvine, California 92697, USA
    J Am Chem Soc 132:13648-50. 2010
    ..We anticipate this affinity selection will be general and become an integral step for the preparation of "plastic antibodies" with near-homogeneous and tailored affinity for target biomacromolecules...
  9. pmc Recognition, neutralization, and clearance of target peptides in the bloodstream of living mice by molecularly imprinted polymer nanoparticles: a plastic antibody
    Yu Hoshino
    Department of Chemistry, University of California, Irvine, California 92697, USA
    J Am Chem Soc 132:6644-5. 2010
    ..Coupled with their biocompatibility and nontoxic characteristics, plastic antibodies offer the potential for neutralizing a wide range of biomacromolecules in vivo...
  10. pmc Synthetic polymer nanoparticles with antibody-like affinity for a hydrophilic peptide
    Zhiyang Zeng
    Department of Chemistry, University of California, Irvine, California 92697, USA
    ACS Nano 4:199-204. 2010
    ..This method can be used for the preparation of nanosized synthetic polymers with antibody-like affinity for hydrophilic peptides and proteins ("plastic antibodies")...
  11. pmc Design of synthetic polymer nanoparticles that capture and neutralize a toxic peptide
    Yu Hoshino
    Department of Chemistry, University of California, Irvine, Irvine, CA 92697, USA
    Small 5:1562-8. 2009
    ..NPs with optimized composition are capable of neutralizing the toxin even in a complex biological milieu. It is anticipated that this strategy will be a starting point for the design of synthetic alternatives to antibodies...