Human Monoclonal Antibody Fragments Targeting Matrilin-3 in Growth Plate Cartilage
Many genetic disorders, including chondrodysplasias, and acquired disorders impair growth plate function, resulting in short and sometimes malformed bones. There are multiple endocrine and paracrine factors that promote chondrogenesis at the growth plate, which could potentially be used to treat these disorders. Targeting these growth factors specifically to the growth plate might augment the therapeutic skeletal effect while diminishing undesirable effects on non-target tissues.
Using yeast display technology, we selected single-chain variable antibody fragments that bound to human and mouse matrilin-3, an extracellular matrix protein specifically expressed in cartilage tissue. The ability of the selected antibody fragments to bind matrilin-3 and to bind cartilage tissue in vitro and in vivo was assessed by ELISA and immunohistochemistry.
We identified antibody fragments that bound matrilin-3 with high affinity and also bound with high tissue specificity to cartilage homogenates and to cartilage structures in mouse embryo sections. When injected intravenously in mice, the antibody fragments specifically homed to cartilage.
Yeast display successfully selected antibody fragments that are able to target cartilage tissue in vivo. Coupling these antibodies to chondrogenic endocrine and paracrine signaling molecules has the potential to open up new pharmacological approaches to treat childhood skeletal growth disorders.
KEY WORDSchildhood growth drug targeting scFv skeletal diseases yeast display
Analysis of variance
Bone morphogenetic protein
Bovine serum albumin
C-type natriuretic peptide
Half maximal effective concentration
Enzyme-linked immunosorbent assay
Fetal calf serum
Horse radish peroxidase
Insulin-like growth factor
Phosphate buffered saline
Single-chain variable fragment
This work was supported by the Intramural Research Programs of the Eunice Kennedy Shriver National Institute of Child Health and Human Development and the National Cancer Institute, National Institutes of Health. C.S.C., Z.Z., J.C.L., D.D. and J.B are co-inventors in a provisional patent application (U.S. Patent Application No. 61/927,904) submitted by the National Institutes of Health.
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