Description | Targets panned | DPIS

The Marasco laboratory at Dana-Farber Cancer Institute (DFCI) has constructed two phage display libraries containing 12 billion (Mehta I) and 15 billion (Mehta II) human sFv antibody phages, which the Center of Therapeutic Antibody Engineering will use to provide NFCR scientists with high affinity human antibodies to support their basic cancer research and/or clinical studies for the treatment of cancer. Description of the Mehta I and II human sFv antibody phage libraries is provided below. Questions related to future commercial licensing of antibodies derived from the Mehta I and II human sFv antibody phage display libraries should be directed to Dr. Wayne A. Marasco.

Construction

To construct the Mehta I and II sFv-phage libraries, we isolated 2.75 x 108 B cells from the peripheral blood of 57 volunteers at DFCI. The initial VH and VL repertoires consisted of 3.5 x 108 VH, 120 x 106 Vkappa and 80 x 106 Vlambda transformants. The Mehta I library was constructed through the random assembly of 2.75 x 108 VH genes with 1.6 x 106 Vkappa and 1.6 x 106 Vlambda genes to yield 12 billion member library. The Mehta II library was constructed identically except that Vkappa genes were not used and a 15 billion member library was obtained.

Characterization

The genetic diversity of the libraries was determined by DNA sequence analysis of unselected sFv-phage from each library. Genetic studies on circa 100 clones have confirmed that each of the major heavy and light chain immunoglobulin germline gene families are represented. These two human sFv libraries are among a handful of the largest human sFv-phage display libraries ever made (Nissim, 1994; Griffiths, 1994; Vaughan, 1996; Sheets, 1998, de Haard, 1999).

Selection

Numerous human sFv antibodies have been isolated from the Mehta libraries. Examples are provided in the next Table.

[ back to top ]