title: Isolation and characterisation of a recombinant antibody fragment that binds NCAM1-expressing intervertebral disc cells.
abstract: Degeneration of the intervertebral discs (IVD) is a leading cause of neck and low back pain. Degeneration begins in the central nucleus pulposus region, leading to loss of IVD osmotic properties. Regeneration approaches include administration of matrix-mimicking scaffolds, cells and/or therapeutic factors. Cell-targeting strategies are likely to improve delivery due to the low cell numbers in the IVD. Single-chain antibody fragments (scFvs) that bind IVD cells were isolated for potential delivery of therapeutics to degenerated IVD. The most cell-distal domain of neural cell adhesion molecule 1 (NCAM1) was cloned and expressed in Escherichia coli. Phage display technology was used to isolate a human scFv against the recombinant domain by panning a scFv library on the immobilised protein. The isolated scFv bound cultured rat astrocytes, as well as bovine nucleus pulposus and annulus fibrosus cells in immunocytochemical studies. The scFv also labelled cells in bovine spinal cord and six-month and two-year old bovine IVD sections by immunohistochemistry. Antibody fragments can provide cell-binding moieties at improved cost, time, yield and functionalisation potential over whole antibodies. The described scFv has potential application in delivery of therapeutics to NCAM1-expressing cells in degenerated IVD.
authors.affiliation: Network of Excellence for Functional Biomaterials (NFB), National University of Ireland, Galway (NUI Galway), Galway, Ireland ; Microbiology, NUI Galway, Galway, Ireland.
authors.forename: Claire
authors.initials: C
authors.lastname: Cunningham
authors.affiliation: Network of Excellence for Functional Biomaterials (NFB), National University of Ireland, Galway (NUI Galway), Galway, Ireland ; Microbiology, NUI Galway, Galway, Ireland.
authors.forename: Akshay
authors.initials: A
authors.lastname: Srivastava
authors.affiliation: Microbiology, NUI Galway, Galway, Ireland.
authors.forename: Estelle
authors.initials: E
authors.lastname: Collin
authors.affiliation: Microbiology, NUI Galway, Galway, Ireland ; AO Research Institute, Davos, Switzerland.
authors.forename: Sibylle
authors.initials: S
authors.lastname: Grad
authors.affiliation: Microbiology, NUI Galway, Galway, Ireland ; AO Research Institute, Davos, Switzerland.
authors.forename: Mauro
authors.initials: M
authors.lastname: Alini
authors.affiliation: Microbiology, NUI Galway, Galway, Ireland.
authors.forename: Abhay
authors.initials: A
authors.lastname: Pandit
authors.affiliation: Network of Excellence for Functional Biomaterials (NFB), National University of Ireland, Galway (NUI Galway), Galway, Ireland ; Microbiology, NUI Galway, Galway, Ireland.
authors.forename: J Gerard
authors.initials: JG
authors.lastname: Wall
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D019002:CD56 Antigen / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D002417:Cattle; D002478:Cells, Cultured; D016503:Drug Delivery Systems; D015870:Gene Expression; D006801:Humans; D007403:Intervertebral Disc / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D055959:Intervertebral Disc Degeneration / Q000188:drug therapy* / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D051379:Mice; D051381:Rats; D057127:Single-Chain Antibodies / Q000276:immunology / Q000494:pharmacology*
chemical_list: D019002:CD56 Antigen; C522918:NCAM1 protein, human; D057127:Single-Chain Antibodies