Phage display of a catalytic antibody to optimize affinity for transition-state analog binding

Catalytic antibodies have shown great promise for catalyzing a tremendously diverse set of natural and unnatural chemical transformations. However, few catalytic antibodies have efficiencies that approach those of natural enzymes. In principle, random mutagenesis procedures such as phage display could be used to improve the catalytic activities of existing antibodies; however, these studies have been hampered by difficulties in the recombinant expression of antibodies. Here, we have grafted the antigen binding loops from a murine-derived catalytic antibody, 17E8, onto a human antibody framework in an effort to overcome difficulties associated with recombinant expression and phage display of this antibody. “Humanized” 17E8 retained similar catalytic and hapten binding properties as the murine antibody while levels of functional Fab displayed on phage were 200-fold higher than for a murine variable region/human constant region chimeric Fab. This construct was used to prepare combinatorial libraries. Affinity panning of these resulted in the selection of variants with 2- to 8-fold improvements in binding affinity for a phosphonate transition-state analog. Surprisingly, none of the affinity-matured variants was more catalytically active than the parent antibody and some were significantly less active. By contrast, a weaker binding variant was identified with 2-fold greater catalytic activity and incorporation of a single substitution (Tyr-100aH → Asn) from this variant into the parent antibody led to a 5-fold increase in catalytic efficiency. Thus, phage display methods can be readily used to optimize binding of catalytic antibodies to transition-state analogs, and when used in conjunction with limited screening for catalysis can identify variants with higher catalytic efficiencies.

The brown-tail moth, Euproctis chrysorrhcea (L.). A report on the life history and habits of the imported brown-tail moth, together with a description of the remedies best suited for destroying it. By Charles H. Fernald ... and Archie H. Kirkland ... Pub. under the direction of the State board of agriculture by authority of the legislature.

1903

The brown-tail moth, Euproctis chrysorrhcea (L.). A report on the life history and habits of the imported brown-tail moth, together with a description of the remedies best suited for destroying it. By Charles H. Fernald ... and Archie H. Kirkland ... Pub. under the direction of the State board of agriculture by authority of the legislature.

1903

The new Soviet legislature : Committee on Defense and State Security : report of the Committee on Armed Services, House of Representatives, One Hundred First Congress, second session.

"The attached report was prepared ... by Dr. Mark Lowenthal ..."--p. iv.

The contribution of library science to efficiency in modern business : outline of two lectures-- Illinois State Library School, May 9-10, 1912.

Mimeographed copy.

The Jalayirids : Dynastic State Formation in the Mongol Middle East /

This book examines the rise and collapse of Mongol rule in Iran and Iraq, and its revival by a family of sultans who claimed to be the rightful heirs to the Mongol khans. The Jalayirids offers a glimpse at a long overlooked but critical period in the history of the Middle East in the late medieval period. This title was made Open Access by libraries from around the world through Knowledge Unlatched.

State of the union message : message from the President of the United States transmitting a report on the state of the union.

"January 23, 2004."

Dental bibliography ... literature of dental science and art as found in the libraries of the New York academy of medicine and Bernhard Wolf Weinberger,

[pt. I.] A reference index.--pt. II. A subject index, with additional reference index.

Federal hazardous waste sites in the state of Hawaii : compliance, cleanup and waste management : hearing before the Committee on Governmental Affairs, United States Senate, One Hundred First Congress, second session, August 20, 1990.

Shipping list no.: 92-0573-P.

Civil service and libraries /

Bibliography: p. 37-40.

Annual report of the Library Commission for the state of Delaware for the fiscal year ...

Title from cover.

Report on the geological survey of the State of Iowa, to the Thirteenth General Assembly, January, 1870, containing results of examinations and observations made within the years 1866, 1867, 1868, and 1869.

Photocopy. Iowa City, Iowa, University of Iowa Libraries, 1980.

New York State Library [annual report] /

From 1889 to 1918 the reports consist of the Report of the director and appendixes, which from 1893 include various bulletins issued by the library (Additions; Bibliography; History; Legislation; Library school; Public libraries) These, including the Report of the director, were each issued also separately.

CATV and its implications for libraries : proceedings of a conference /

Includes bibliographical references and index.

The use of computers in literature searching and related reference activities in libraries : papers presented at the 1975 Clinic on Library Applications of Data Processing, April 27-30, 1975 /

Includes bibliographical references.