Document Detail

A combinatorial approach for directing the amount of fibronectin fibrils assembled by cells that uses surfaces derivatized with mixtures of fibronectin and cell binding domains.
MedLine Citation:
PMID:  22467639     Owner:  NLM     Status:  MEDLINE    
Fibrillar fibronectin (FN) has the crucial role of attracting and attaching cells as well as molecules that mediate tissue repair during wound healing. A previous study demonstrated higher extracellular staining of FN fibrils in cells cultured on surfaces tethered with an equimolar mixture of a FN binding domain and FN's cell binding domain, III1-2 and III9-10 respectively, than on surfaces with III9-10 alone. The effect of varying surface amounts of III1-2 and III9-10 on the quantity of FN fibrils formed by NIH-3T3 fibroblasts was examined. GST tagged III1-2 and III9-10 were conjugated to polyurethane surfaces and ELISAs were used to identify the experimental design space or the range of concentrations of GST-III1-2 and GST-III9-10 that demarcated the limits of protein loading on the surface. When GST-III1-2 was fixed and GST-III9-10 varied within the design space, the amount of FN fibrils measured by immunoblotting detergent insoluble cell lysates was dependent on the ratio of III9-10 to III1-2. When the total protein concentration was fixed and the mixture composition of GST-III1-2 and GST-III9-10 varied such that it optimally covered the design space, a parabolic relationship between FN fibril amount and the ratio of III9-10 to III1-2 was obtained. This relationship had a maximum value when the surface was bonded to equal amounts of III1-2 and III9-10 (P<0.05). Thus the ratio of III9-10 to III1-2 can be utilized to direct the quantity of FN fibrils formed on surfaces.
Pradnya P Kshatriya; Stella W Karuri; Chunyi Chiang; Nancy W Karuri
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Publication Detail:
Type:  Journal Article     Date:  2012-05-21
Journal Detail:
Title:  Biotechnology progress     Volume:  28     ISSN:  1520-6033     ISO Abbreviation:  Biotechnol. Prog.     Publication Date:    2012 May-Jun
Date Detail:
Created Date:  2012-06-11     Completed Date:  2012-11-05     Revised Date:  2012-12-06    
Medline Journal Info:
Nlm Unique ID:  8506292     Medline TA:  Biotechnol Prog     Country:  United States    
Other Details:
Languages:  eng     Pagination:  862-71     Citation Subset:  IM    
Copyright Information:
Copyright © 2012 American Institute of Chemical Engineers (AIChE).
Department of Chemical and Biological Engineering, Illinois Institute of Technology, Chicago, IL 60616, USA.
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MeSH Terms
Cells, Cultured
Combinatorial Chemistry Techniques*
Enzyme-Linked Immunosorbent Assay
Fibronectins / metabolism*
Glutathione Transferase / metabolism
NIH 3T3 Cells
Protein Binding
Reg. No./Substance:
0/Fibronectins; EC Transferase

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