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Generation of a polyclonal fab phage display library to the human breast carcinoma cell line BT-20.

Publication ,  Journal Article
Santora, KE; Sarantopoulos, S; Den, W; Petersen-Mahrt, S; Sompuram, SR; Sharon, J
Published in: Comb Chem High Throughput Screen
February 2000

We have previously described a vector system for generating recombinant polyclonal antibody libraries. This system uses bidirectional phagemid and mammalian expression vectors to facilitate mass transfer of selected variable light and variable heavy (VL-VH) region gene pairs from the phagemid to the mammalian vector, to express polyclonal libraries of whole IgG antibodies. We report here the first stage of generating a polyclonal antibody library to the human breast carcinoma cell line BT-20, using this vector system. VL and VH region gene pairs were obtained from a mouse immunized with BT-20 cells, and cloned, in opposite transcriptional orientations, in the bidirectional phagemid vector, to produce an Fab phage display library. This library was selected by panning on BT-20 cells and shown to bind specifically to BT-20 cells. Such libraries, after suitable negative selection to eliminate major reactivities against normal tissue, could be transferred in mass to our bidirectional mammalian expression vector for production of libraries of chimeric antibodies with mouse V regions and human constant (C) regions. These polyclonal antibody libraries will mediate effector functions and are expected to be useful for breast cancer therapy, as well as diagnosis.

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Published In

Comb Chem High Throughput Screen

DOI

ISSN

1386-2073

Publication Date

February 2000

Volume

3

Issue

1

Start / End Page

51 / 57

Location

United Arab Emirates

Related Subject Headings

  • Tumor Cells, Cultured
  • Sequence Homology, Nucleic Acid
  • Organic Chemistry
  • Molecular Sequence Data
  • Mice, Inbred BALB C
  • Mice
  • Immunoglobulin Fab Fragments
  • Humans
  • Female
  • DNA, Complementary
 

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Santora, K. E., Sarantopoulos, S., Den, W., Petersen-Mahrt, S., Sompuram, S. R., & Sharon, J. (2000). Generation of a polyclonal fab phage display library to the human breast carcinoma cell line BT-20. Comb Chem High Throughput Screen, 3(1), 51–57. https://doi.org/10.2174/1386207003327765
Santora, K. E., S. Sarantopoulos, W. Den, S. Petersen-Mahrt, S. R. Sompuram, and J. Sharon. “Generation of a polyclonal fab phage display library to the human breast carcinoma cell line BT-20.Comb Chem High Throughput Screen 3, no. 1 (February 2000): 51–57. https://doi.org/10.2174/1386207003327765.
Santora KE, Sarantopoulos S, Den W, Petersen-Mahrt S, Sompuram SR, Sharon J. Generation of a polyclonal fab phage display library to the human breast carcinoma cell line BT-20. Comb Chem High Throughput Screen. 2000 Feb;3(1):51–7.
Santora, K. E., et al. “Generation of a polyclonal fab phage display library to the human breast carcinoma cell line BT-20.Comb Chem High Throughput Screen, vol. 3, no. 1, Feb. 2000, pp. 51–57. Pubmed, doi:10.2174/1386207003327765.
Santora KE, Sarantopoulos S, Den W, Petersen-Mahrt S, Sompuram SR, Sharon J. Generation of a polyclonal fab phage display library to the human breast carcinoma cell line BT-20. Comb Chem High Throughput Screen. 2000 Feb;3(1):51–57.
Journal cover image

Published In

Comb Chem High Throughput Screen

DOI

ISSN

1386-2073

Publication Date

February 2000

Volume

3

Issue

1

Start / End Page

51 / 57

Location

United Arab Emirates

Related Subject Headings

  • Tumor Cells, Cultured
  • Sequence Homology, Nucleic Acid
  • Organic Chemistry
  • Molecular Sequence Data
  • Mice, Inbred BALB C
  • Mice
  • Immunoglobulin Fab Fragments
  • Humans
  • Female
  • DNA, Complementary