Antibody fragments directed against different portions of the human neural cell adhesion molecule L1 act as inhibitors or activators of L1 function.
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Antibody fragments directed against different portions of the human neural cell adhesion molecule L1 act as inhibitors or activators of L1 function. / Wang, Yan; Loers, Gabriele; Pan, Hong-Chao; Gouveia, Ricardo; Zhao, Wei-Jiang; Shen, Yan-Qin; Kleene, Ralf; Costa, Julia; Schachner, Melitta.
in: PLOS ONE, Jahrgang 7, Nr. 12, 12, 2012, S. 52404.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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T1 - Antibody fragments directed against different portions of the human neural cell adhesion molecule L1 act as inhibitors or activators of L1 function.
AU - Wang, Yan
AU - Loers, Gabriele
AU - Pan, Hong-Chao
AU - Gouveia, Ricardo
AU - Zhao, Wei-Jiang
AU - Shen, Yan-Qin
AU - Kleene, Ralf
AU - Costa, Julia
AU - Schachner, Melitta
PY - 2012
Y1 - 2012
N2 - The neural cell adhesion molecule L1 plays important roles in neuronal migration and survival, neuritogenesis and synaptogenesis. L1 has also been found in tumors of different origins, with levels of L1 expression correlating positively with the metastatic potential of tumors. To select antibodies targeting the varied functions of L1, we screened the Tomlinson library of recombinant human antibody fragments to identify antibodies binding to recombinant human L1 protein comprising the entire extracellular domain of human L1. We obtained four L1 binding single-chain variable fragment antibodies (scFvs), named I4, I6, I13, and I27 and showed by enzyme-linked immunosorbent assay (ELISA) that scFvs I4 and I6 have high affinity to the immunoglobulin-like (Ig) domains 1-4 of L1, while scFvs I13 and I27 bind strongly to the fibronectin type III homologous (Fn) domains 1-3 of L1. Application of scFvs I4 and I6 to human SK-N-SH neuroblastoma cells reduced proliferation and transmigration of these cells. Treatment of SK-N-SH cells with scFvs I13 and I27 enhanced cell proliferation and migration, neurite outgrowth, and protected against the toxic effects of H(2)O(2) by increasing the ratio of Bcl-2/Bax. In addition, scFvs I4 and I6 inhibited and scFvs I13 and I27 promoted phosphorylation of src and Erk. Our findings indicate that scFvs reacting with the immunoglobulin-like domains 1-4 inhibit L1 functions, whereas scFvs interacting with the fibronectin type III domains 1-3 trigger L1 functions of cultured neuroblastoma cells.
AB - The neural cell adhesion molecule L1 plays important roles in neuronal migration and survival, neuritogenesis and synaptogenesis. L1 has also been found in tumors of different origins, with levels of L1 expression correlating positively with the metastatic potential of tumors. To select antibodies targeting the varied functions of L1, we screened the Tomlinson library of recombinant human antibody fragments to identify antibodies binding to recombinant human L1 protein comprising the entire extracellular domain of human L1. We obtained four L1 binding single-chain variable fragment antibodies (scFvs), named I4, I6, I13, and I27 and showed by enzyme-linked immunosorbent assay (ELISA) that scFvs I4 and I6 have high affinity to the immunoglobulin-like (Ig) domains 1-4 of L1, while scFvs I13 and I27 bind strongly to the fibronectin type III homologous (Fn) domains 1-3 of L1. Application of scFvs I4 and I6 to human SK-N-SH neuroblastoma cells reduced proliferation and transmigration of these cells. Treatment of SK-N-SH cells with scFvs I13 and I27 enhanced cell proliferation and migration, neurite outgrowth, and protected against the toxic effects of H(2)O(2) by increasing the ratio of Bcl-2/Bax. In addition, scFvs I4 and I6 inhibited and scFvs I13 and I27 promoted phosphorylation of src and Erk. Our findings indicate that scFvs reacting with the immunoglobulin-like domains 1-4 inhibit L1 functions, whereas scFvs interacting with the fibronectin type III domains 1-3 trigger L1 functions of cultured neuroblastoma cells.
KW - Humans
KW - Cell Line
KW - Protein Binding
KW - Cell Proliferation/drug effects
KW - Cell Movement/drug effects
KW - Phosphorylation/drug effects
KW - Hydrogen Peroxide/toxicity
KW - Neural Cell Adhesion Molecule L1/agonists/antagonists & inhibitors/metabolism
KW - Neurites/drug effects
KW - Protein Interaction Domains and Motifs
KW - Recombinant Proteins/isolation & purification/metabolism/pharmacology
KW - Single-Chain Antibodies/isolation & purification/metabolism/pharmacology
KW - src-Family Kinases/metabolism
KW - Humans
KW - Cell Line
KW - Protein Binding
KW - Cell Proliferation/drug effects
KW - Cell Movement/drug effects
KW - Phosphorylation/drug effects
KW - Hydrogen Peroxide/toxicity
KW - Neural Cell Adhesion Molecule L1/agonists/antagonists & inhibitors/metabolism
KW - Neurites/drug effects
KW - Protein Interaction Domains and Motifs
KW - Recombinant Proteins/isolation & purification/metabolism/pharmacology
KW - Single-Chain Antibodies/isolation & purification/metabolism/pharmacology
KW - src-Family Kinases/metabolism
U2 - 10.1371/journal.pone.0052404
DO - 10.1371/journal.pone.0052404
M3 - SCORING: Journal article
VL - 7
SP - 52404
JO - PLOS ONE
JF - PLOS ONE
SN - 1932-6203
IS - 12
M1 - 12
ER -
