Synthesis and characterization of CN-modified protein analogues as potential vibrational contrast agents.
A recombinant VH single-domain antibody recognizing staphylococcal protein A was functionalized on reactive lysine residues with N-hydroxysuccimidyl-activated 4-cyanobenzoate. Surface plasmon resonance analysis of antibody-antigen binding revealed that modified and unmodified antibodies bound protein A with similar affinities. Raman imaging of the modified antibodies indicated that the benzonitrile group provides vibrational contrast enhancement in a region of the electromagnetic spectrum that is transparent to cellular materials. Thus, the modified single-domain antibody may be amenable to detecting protein A from samples of the human pathogen Staphylococcus aureus using vibronic detection schemes such as Raman and coherent anti-Stokes Raman scattering. The generality of this labeling strategy should make it applicable to modifying an array of proteins with varied structure and function.
Noestheden
Matthew
M
Steacie Institute for Molecular Sciences, National Research Council of Canada, Ottawa, Canada.
Hu
Qingyan
Q
Tay
Li-Lin
LL
Tonary
Angela M
AM
Stolow
Albert
A
MacKenzie
Roger
R
Tanha
Jamshid
J
Pezacki
John Paul
JP
D000937:Antigen-Antibody Reactions; D001565:Benzoates / Q000138:chemical synthesis* / Q000737:chemistry* / Q000494:pharmacology; D015394:Molecular Structure; D017434:Protein Structure, Tertiary; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000187:drug effects; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013059:Spectrum Analysis, Raman / Q000379:methods; D013205:Staphylococcal Protein A / Q000737:chemistry* / Q000187:drug effects; D013211:Staphylococcus aureus / Q000737:chemistry / Q000187:drug effects; D013329:Structure-Activity Relationship; D013388:Succinimides / Q000737:chemistry; D020349:Surface Plasmon Resonance / Q000379:methods; D014732:Vibration
D001565:Benzoates; D013205:Staphylococcal Protein A; D013388:Succinimides; D012710:Serum Albumin, Bovine; C097383:4-cyanobenzoic acid; C001426:N-hydroxysuccinimide