W. Alton Jones Cell Science Center Incorporated

J Denry Sato

Lihua Wang

Tetsuji Okamoto

Dianging Wu

Yoshinari Myoken

Hybridomas producing monoclonal antibodies that specifically bind to the human EGF receptor and compete with EGF for binding to the EGF receptor are described. The subject monoclonal antibodies bind to an epitope located between residues Ala-351 and Asp-364 of the EGF receptor, which is not denatured by boiling, treatment with detergent or treatment with a reducing agent.

Hybridomas producing monoclonal antibodies which specifically bind to

Cholesterol auxotrophy of myeloma cells is used as the basis for selecting hybridomas. The outgrowth of nascent hybridomas in a cholesterol-free medium was 3- to 9-fold more efficient than that in HAT medium and resulted in 3- to 13-times as many antigen-reactive hybridoma cells. This method of hybridoma selection can be applied with any sterol-dependent parent cell line. The nutrient medium is also preferably free of Ham's F-12 nutrient mixture.

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Lake Placid, NY, United States of America

J Denry Sato