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A High Throughput Enzyme-Linked Immunosorbent Assay for Inhibitors of the Interaction Between Retinoblastoma Protein and the Leu-X-Cys-X-Glu Motif

Undergraduate student on placement from the Department of Biochemistry, University of Bath, Bath BA2 7AY, United Kingdom

Al Peng Teoh

Centre for Natural Product Research, Institute of Molecular and Cell Biology, National University of Singapore, 15 Lower Kent Ridge Road, Singapore 119076

Arasu Ganesan

Centre for Natural Product Research, Institute of Molecular and Cell Biology, National University of Singapore, 15 Lower Kent Ridge Road, Singapore 119076

A 96-well enzyme-linked immunosorbent assay was developed to discover compounds that inhibit the binding of the Leu-X-Cys-X-Glu (LXCXE) motif to the retinoblastoma tumor suppressor protein (pRB). The assay uses a LXCXE-containing multiple antigenic peptide (MAP) which is immobilized on a microtiter plate. A truncated form of pRB is added and the amount bound detected by a monoclonal antibody. This rapid assay was employed in high throughput screening of crude natural product extracts and discrete compounds.

Journal of Biomolecular Screening, Vol. 2, No. 4, 207-211 (1997)
DOI: 10.1177/108705719700200405


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