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.
Department of Pathology, University Medical Center Utrecht, PO Box 85500, 3508 GA Utrecht, The Netherlands.
Arjan J
AJ
Groot
Mohamed
M
El Khattabi
Norman
N
Sachs
Petra
P
van der Groep
Elsken
E
van der Wall
Paul J
PJ
van Diest
Arnoud
A
Sonnenberg
C Theo
CT
Verrips
Marc
M
Vooijs
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