BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 577

Peptides:

Sequences	Affinity Values
GEPQTKLFSFPL(10)	[1.13 ± 0.14]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (subtractive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 19103307

Target Name:

Anti-CTGF monoclonal antibody 7G2

Template Name:

Connective tissue growth factor

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

Ph.D.-12 phage display library (X12)

Affinity Measurement Method:

ELISA

Affinity Measurement Description:

Microtiter wells were coated with 2 μg/ml of ZD521 peptide, TrxA-ZD521, or TrxA and blocked with PBS containing 10% fetal bovine sera. 7G2(anti-CTGF antibody) in the concentrations of 0, 0.1, 0.5, 1, 2 μg/ml was added to the wells. Bound 7G2 was detected with HRP-conjugated anti-mouse IgG antibody and then with the substrate. The values are recorded at OD450 nm. SDs are indicated by error bars. Only the value of the binding of ZD521 peptide to 7G2 in the concentration of 2 μg/ml was shown. Data were reproduced from the graph.

Brief Description:

This monoclonal antibody was produced by the hybridoma technique using recombinant CTGF C-terminal domain as the immunogen. The results of ELISA assays and Westen-blot showed that 7G2 could specifically bind to TrxA-CTGF/C and kidney mesangial cell lysates, but not to TrxA. The phage library was reabsorbed by mouse serum IgG and BSA to remove non-specific binders, and then specifically absorbed with 7G2.

BiopanningDataSet Comments:

After 3 rounds of selection, roughly 31.3% (10/32) of the phage clones analyzed exhibited 7G2 binding activity by ELISA. All 10 positive phage clones were sequenced, and the sequences of all clones were identical, i.e. GEPQTKLFSFPL. The antiserum from mice immunized with TrxA-GEPQTKLFSFPL could also bind to CTGF/C recombinant protein and native CTGF, as well as significantly inhibit the proliferation of kidney mesangial cells induced by CTGF/C.

Curator:

Curation Date:

0000-00-00

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