TY - JOUR
T1 - Single-molecule selection and recovery of structure-specific antibodies using atomic force microscopy
AU - Shlyakhtenko, Luda S.
AU - Yuan, Bin
AU - Emadi, Sharareh
AU - Lyubchenko, Yuri L.
AU - Sierks, Michael
N1 - Funding Information:
This work was supported in part by grants from the National Institutes of Health (1PN1EY016593 and 5R01GM062235), MJ Fox Foundation, and Arizona Biomedical Research Commission.
PY - 2007/9
Y1 - 2007/9
N2 - Protein misfolding and aggregation are a common thread in numerous diseases including Alzheimer's, Parkinson's, Huntington's, amyotrophic lateral sclerosis, diabetes, and prion-related diseases. Elucidation of the role played by the various protein forms in these diseases requires reagents that can target specific protein forms. Here we present a method to isolate antibodies that bind to a specific protein form. We combined the imaging and nanomanipulation capabilities of atomic force microscopy (AFM) with the protein diversity of phage display antibody libraries to develop a technology that allows us to recover a single antibody molecule that is bound to a single protein molecular target. The target protein-antibody complex is first imaged by AFM, the AFM tip is then manipulated by nanolithography over the target antibody to recover the associated phage, and the antibody gene is recovered from the single phage particle by polymerase chain reaction.
AB - Protein misfolding and aggregation are a common thread in numerous diseases including Alzheimer's, Parkinson's, Huntington's, amyotrophic lateral sclerosis, diabetes, and prion-related diseases. Elucidation of the role played by the various protein forms in these diseases requires reagents that can target specific protein forms. Here we present a method to isolate antibodies that bind to a specific protein form. We combined the imaging and nanomanipulation capabilities of atomic force microscopy (AFM) with the protein diversity of phage display antibody libraries to develop a technology that allows us to recover a single antibody molecule that is bound to a single protein molecular target. The target protein-antibody complex is first imaged by AFM, the AFM tip is then manipulated by nanolithography over the target antibody to recover the associated phage, and the antibody gene is recovered from the single phage particle by polymerase chain reaction.
KW - Atomic force microscopy
KW - Phage display
KW - Protein misfolding
KW - Single molecule
KW - Single-chain antibody fragment
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U2 - 10.1016/j.nano.2007.06.001
DO - 10.1016/j.nano.2007.06.001
M3 - Article
C2 - 17662669
AN - SCOPUS:34548248155
VL - 3
SP - 192
EP - 197
JO - Nanomedicine: Nanotechnology, Biology, and Medicine
JF - Nanomedicine: Nanotechnology, Biology, and Medicine
SN - 1549-9634
IS - 3
ER -