TY - JOUR
T1 - Efficient method to optimize antibodies using avian leukosis virus display and eukaryotic cells
AU - Yu, Changming
AU - Pike, Gennett M.
AU - Rinkoski, Tommy A.
AU - Correia, Cristina
AU - Kaufmann, Scott H.
AU - Federspiel, Mark J.
AU - Weiss, Robin A.
PY - 2015/8/11
Y1 - 2015/8/11
N2 - Antibody-based therapeutics have now had success in the clinic. The affinity and specificity of the antibody for the target ligand determines the specificity of therapeutic delivery and off-target side effects. The discovery and optimization of high-affinity antibodies to important therapeutic targets could be significantly improved by the availability of a robust, eukaryotic display technology comparable to phage display that would overcome the protein translation limitations of microorganisms. The use of eukaryotic cells would improve the diversity of the displayed antibodies that can be screened and optimized as well as more seamlessly transition into a large-scale mammalian expression system for clinical production. In this study, we demonstrate that the replication and polypeptide display characteristics of a eukaryotic retrovirus, avian leukosis virus (ALV), offers a robust, eukaryotic version of bacteriophage display. The binding affinity of a model single-chain Fv antibody was optimized by using ALV display, improving affinity >2,000-fold, from micromolar to picomolar levels. We believe ALV display provides an extension to antibody display on microorganisms and offers virus and cell display platforms in a eukaryotic expression system. ALV display should enable an improvement in the diversity of properly processed and functional antibody variants that can be screened and affinity-optimized to improve promising antibody candidates.
AB - Antibody-based therapeutics have now had success in the clinic. The affinity and specificity of the antibody for the target ligand determines the specificity of therapeutic delivery and off-target side effects. The discovery and optimization of high-affinity antibodies to important therapeutic targets could be significantly improved by the availability of a robust, eukaryotic display technology comparable to phage display that would overcome the protein translation limitations of microorganisms. The use of eukaryotic cells would improve the diversity of the displayed antibodies that can be screened and optimized as well as more seamlessly transition into a large-scale mammalian expression system for clinical production. In this study, we demonstrate that the replication and polypeptide display characteristics of a eukaryotic retrovirus, avian leukosis virus (ALV), offers a robust, eukaryotic version of bacteriophage display. The binding affinity of a model single-chain Fv antibody was optimized by using ALV display, improving affinity >2,000-fold, from micromolar to picomolar levels. We believe ALV display provides an extension to antibody display on microorganisms and offers virus and cell display platforms in a eukaryotic expression system. ALV display should enable an improvement in the diversity of properly processed and functional antibody variants that can be screened and affinity-optimized to improve promising antibody candidates.
KW - Antibody binding affinity
KW - Antibody engineering
KW - Avian leukosis virus
KW - Avian leukosis virus polypeptide display
KW - Protein display technology
UR - http://www.scopus.com/inward/record.url?scp=84938947389&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84938947389&partnerID=8YFLogxK
U2 - 10.1073/pnas.1414754112
DO - 10.1073/pnas.1414754112
M3 - Article
C2 - 26216971
AN - SCOPUS:84938947389
SN - 0027-8424
VL - 112
SP - 9860
EP - 9865
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 32
ER -