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Affinity selection of cDNA libraries by lambda phage surface display.

M Niwa¹, H Maruyama, T Fujimoto

¹Department of Cell Biology, The Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA 92037, USA.

|October 31, 2000

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Researchers used bacteriophage lambda surface display to identify novel autoantigens in Sjögren

Area of Science:

Immunology
Molecular Biology
Biotechnology

Background:

Sjögren's syndrome (SS) is an autoimmune disease characterized by autoantibodies targeting self-antigens.
Identifying these autoantigens is crucial for understanding disease mechanisms and developing diagnostics.

Purpose of the Study:

To employ bacteriophage lambda surface display for the isolation of cDNA clones encoding autoantigens recognized by SS patient sera.
To identify novel autoantigens associated with Sjögren's syndrome.

Main Methods:

Construction of cDNA libraries from human HeLa and HepG2 cells using the lambdafoo expression vector.
Repeated affinity selection of these libraries using immobilized SS patient sera.
Isolation and characterization of cDNA clones encoding autoantigens.

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Main Results:

Successful isolation of three novel autoantigen clones and four previously known SS autoantigen clones.
Newly identified autoantigens include the TRK-fused gene product (TFG), survival motor neuron gene product (SMN), and pM5.
pM5 exhibits similarity to the metal-binding domain of human fibroblast collagenase.

Conclusions:

Bacteriophage lambda surface display is a powerful and effective technique for isolating cDNA clones via affinity screening.
This method facilitates the discovery of novel autoantigens, advancing Sjögren's syndrome research.