Reverse proteomic antibody screening identifies anti adhesive VHH targeting VLA-3.
Therapeutic approaches aimed at targeting tumor surface markers using monoclonal antibodies provide a powerful strategy in cancer treatment. Here we report selection of single variable domains (VHH) of llama heavy chain antibodies, using a VHH-phage-display library. A reverse proteomic approach was used to identify the cognate proteins recognized by enriched VHH on HeLa cells. One of these VHH bound the integrin alpha 3 beta 1 (VLA-3) and was further characterized. Most interestingly, this VHH could inhibit VLA-3 mediated cell-matrix adhesion. Our approach provides a fast and efficient method to screen for novel cell surface markers on normal and tumor cells that may find diagnostic or therapeutic application in disease management or treatment.
Groot
Arjan J
AJ
Department of Pathology, University Medical Center Utrecht, PO Box 85500, 3508 GA Utrecht, The Netherlands.
El Khattabi
Mohamed
M
Sachs
Norman
N
van der Groep
Petra
P
van der Wall
Elsken
E
van Diest
Paul J
PJ
Sonnenberg
Arnoud
A
Verrips
C Theo
CT
Vooijs
Marc
M
D000268:Adhesiveness; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000032:analysis*; D002161:Camelids, New World; D002448:Cell Adhesion; D002462:Cell Membrane / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005109:Extracellular Matrix / Q000276:immunology; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000737:chemistry / Q000276:immunology*; D047468:Immunoprecipitation; D039201:Integrin alpha3beta1 / Q000737:chemistry / Q000276:immunology*; D015603:Keratinocytes / Q000166:cytology / Q000276:immunology; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results
D000906:Antibodies; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D039201:Integrin alpha3beta1