title: Single chain fragment variable antibodies developed by using as target the 3rd fibronectin type III homologous repeat fragment of human neural cell adhesion molecule L1 promote cell migration and neuritogenesis.
abstract: L1CAM plays important roles during ontogeny, including promotion of neuronal cell migration and neuritogenesis, and stimulation of axonal outgrowth, fasciculation and myelination. These functions are at least partially exerted through a 16-mer amino acid sequence in the third fibronectin type III-like repeat of L1, which associates with several interaction partners, including integrins, other adhesion molecules and growth factor receptors. Here, using the Tomlinson I library for phage display, we obtained two single-chain variable fragment antibodies (scFvs) against this peptide sequence of human L1, hereafter called H3 peptide. Both scFvs recognize the H3 peptide and the extracellular domain of L1, as tested by enzyme-linked immunosorbent assay (ELISA), Western blot analysis and immunofluorescence staining of L1 expresssing cells. Furthermore, both scFvs reduce U-87 MG cell adhesion to fibronectin, while stimulating cell migration. Application of scFvs to human neuroblastoma SK-N-SH cells promote process outgrowth. Similar to triggering of endogenous L1 functions at the cell surface, both scFvs activate the signal transducers Erk and Src in these cells. Our results indicate that scFvs against a functionally pivotal domain in L1 trigger its regeneration-beneficial functions in vitro, encouraging thoughts on therapy of neurodegenerative diseases in the hope to ameliorate human nervous system diseases.
authors.affiliation: Center for Neuroscience, Shantou University Medical College, 22 Xin Ling Road, Jinping District, Shantou, Guangdong 515041, People׳s Republic of China.
authors.forename: Dan-Yang
authors.initials: DY
authors.lastname: Tang
authors.affiliation: Center for Neuroscience, Shantou University Medical College, 22 Xin Ling Road, Jinping District, Shantou, Guangdong 515041, People׳s Republic of China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Center for Neuroscience, Shantou University Medical College, 22 Xin Ling Road, Jinping District, Shantou, Guangdong 515041, People׳s Republic of China.
authors.forename: Xuan-Jun
authors.initials: XJ
authors.lastname: Zhao
authors.affiliation: Center for Neuroscience, Shantou University Medical College, 22 Xin Ling Road, Jinping District, Shantou, Guangdong 515041, People׳s Republic of China. Electronic address: schachner@stu.edu.cn.
authors.forename: Melitta
authors.initials: M
authors.lastname: Schachner
authors.affiliation: Center for Neuroscience, Shantou University Medical College, 22 Xin Ling Road, Jinping District, Shantou, Guangdong 515041, People׳s Republic of China. Electronic address: neuromancn@aliyun.com.
authors.forename: Wei-Jiang
authors.initials: WJ
authors.lastname: Zhao
mesh_terms: D001483:Base Sequence; D002448:Cell Adhesion / Q000502:physiology; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology; D061505:Cell Surface Display Techniques; D004789:Enzyme Activation; D004797:Enzyme-Linked Immunosorbent Assay; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005353:Fibronectins / Q000276:immunology*; D006801:Humans; D039842:Neural Cell Adhesion Molecule L1 / Q000276:immunology*; D016501:Neurites / Q000502:physiology*; D019151:Peptide Library; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D015398:Signal Transduction / Q000276:immunology; D057127:Single-Chain Antibodies / Q000276:immunology*; D019061:src-Family Kinases / Q000378:metabolism
chemical_list: D005353:Fibronectins; D039842:Neural Cell Adhesion Molecule L1; D019151:Peptide Library; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; C501364:fibronectin type III like peptide, human; D019061:src-Family Kinases; D048049:Extracellular Signal-Regulated MAP Kinases
keywords: Cell adhesion molecule L1 (L1CAM, L1); Glioblastoma; Neuroblastoma; Phage display; Single-chain fragment variable antibody (scFv)