National Institutes of Health, a Component of the US Dept. of Health & Human Services

Novartis Ag

Scott & White Healthcare

Jung-Hee Woo

David M Neville

Huaizhong Hu

Jerry Todd Thompson

Shenglin Ma

Mary Ellen Digan

Jonathan Mark Hexham

Yuan-Yi Liu

Arthur E Frankel

Provided are methods and compositions related to diphtheria toxin diabody immunotoxins.

Fold-back diabody diphtheria toxin immunotoxin and methods of use

The present invention described and shown in the specification and drawings provides novel recombinant DT-based immunotoxins, and, more specifically anti-T cell immunotoxin fusion proteins. Also provided are immunotoxins that can be expressed in bacterial, yeast, or mammalian cells. The invention also provides means for expression of the immunotoxin fusion protein. It is emphasized that this abstract is provided to comply with the rules requiring an abstract that will allow a searcher or other reader to quickly ascertain the subject matter of the technical disclosure. It is submitted with the understanding that it will not be used to interpret or limit the scope or meaning of the claims.

Immunotoxin fusion proteins and means for expression thereof

The present invention relates to a method of expressing an immunotoxin instrain mutated to toxin resistance comprising a) growing thein a growth medium comprising an enzymatic digest of protein and yeast extract and maintaining a dissolved oxygen concentration at 40% and above; and b) performing methanol induction with a limited methanol feed of 0.5-0.75 ml/min/IO L of initial volume during induction along with a continuous infusion of yeast extract at a temperature below 17.5&#xb0; C., antifoaming agent supplied up to 0.07%, agitation reduced to 400 RPM, and the induction phase extended out to 163 h.

Methods for expression and purification of immunotoxins

The present invention described and shown in the specification and drawings provides novel recombinant DT-based immunotoxins, and, more specifically anti-T cell immunotoxin fusion proteins. Also provided are immunotoxins that can be expressed in bacterial, yeast, or mammalian cells. The invention also provides means for expression of the immunotoxin fusion protein.

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Rockville, MD, United States of America

Jung-Hee Woo