Xoma Corporation

University of California

Gary L Wilcox

Marc D Better

Randy R Robinson

Shau-Ping Lei

Alvin Y Liu

Randolph Wall

Arnold H Horwitz

Jar-How Lee

Jiunu S Lai

Dan S Ray

Yun-Long Lin

Donald A Kaplan

Arnold H Horowitz

Patrick E Clarke

Modular assembly of antibody genes, antibodies prepared thereby and use

The invention is directed to the pectate lyase B secretion signal and its use to express operably linked sequences in bacterial hosts.

Pectate lyase signal sequence

The invention relates to cDNA genetic sequences, vehicles containing same as well as hosts transformed therewith, for the production of chimeric immunoglobulin molecules, functional fragments thereof and immunoglobulin derivatives exhibiting novel functional properties comprising human constant region modules and non-human variable region modules, or for class switching antibody molecules and/or chains. The invention also relates to DNA coding for pectate lyase signal peptide has been cloned on a plasmid to create a secretion vector which is capable of producing a chosen protein which is transported across the bacterial membrane. The secretion vector has been used to secrete extracellular thaumatin and extracellular chimeric antibody fragments. The proteins produced by this vector have biological activity. The thaumatin is properly folded and the antibody fragments are capable of binding antigens on target cancer cells. The invention also relates to the secretion of chimeric antibodies and fragments thereof from yeast in functional form.

The invention relates to the secretion of heavy chain immunoglobulin fragments and light chain immunoglobulins from prokaryotic hosts using a prokaryotic secretion signal peptide wherein the heavy chain fragments and light chains are capable of associating to form an antigen binding antibody fragment.

The invention relates to cDNA genetic sequences, vehicles containing same as well as hosts transformed therewith, for the production of chimeric immunoglobulin molecules, functional fragments thereof and immunoglobulin derivatives exhibiting novel functional properties comprising human constant region modules and non-human variable region modules, or for class switching antibody molecules and/or chains. The invention also relates to DNA coding for pectate lyase signal peptide has been cloned on a plasmid to create a secretion vector which is capable of producing a chosen protein which is transported across the bacterial membrane. The secretion vector has been used to secrete extracellular thaumatin and extracellular chimeric antibody fragments. The proteins produced by this vector have biological activity. The thaumatin is properly folded and the antibody fragments are capable-of binding antigens on target cancer cells. The invention also relates to the secretion of chimeric antibodies and fragments thereof from yeast in functional form.

The invention relates to DNA coding for a pectate lyase signal peptide which has been inserted into plasmids to create secretion vectors which are capable of producing a chosen protein by directing transport across the bacterial membrane. The secretion vectors have been used to secrete extracellular thaumatin and extracellular antibody fragments. The proteins produced by these vectors have biological activity. The thaumatin is properly folded and the antibody fragments are capable of binding antigens on target cancer cells.

Vectors with pectate lyase signal sequence

Cecropin fusion proteins with suppressed bactericidal activity are provided. The fusion proteins of the invention provide the sequence of a cecropin and can be expressed in a cecropin-sensitive bacterium.

Cecropins fusion proteins

A polynucleotide molecule expressible in a given host comprising the sequence of the araB promoter operably linked to a gene which is heterologous to said host. The heterologous gene codes for a peptide that is biologically active. The invention also relates to a genetic construct which comprises a first genetic sequence coding for cecropin operably linked to a second genetic sequence coding for a polypeptide which is capable of suppressing the bactericidal effect of the resulting fusion protein towards an otherwise cecropin sensitive bacterium.

Method of producing cecropins by microbiological techniques

AraB promoters and method of producing polypeptides, including

A method for providing an image of zones of ultraviolet absorbing material in a medium substantially transparent to ultraviolet light, such as nucleic acid in a polyacrylamide gel, comprises the steps of placing in juxtaposition to the gel standard photographic film and directing ultraviolet light to the gel such that the photographic film is in the path of light exiting the gel. The zones in the gel absorb one or more discrete wavelengths of ultraviolet light, and the film is exposed by incident ultraviolet light such that a photographic image of the nucleic acid zones is created on the photographic film. Apparatus for providing ultraviolet images of ultraviolet light absorbing material in a medium relatively transparent to ultraviolet light comprises an enclosure in which a medium with zones of ultraviolet light absorbing material is placed. The closure contains an aperture for reception of light from an ultraviolet light source and is adapted to accommodate a strip of photographic film juxtaposed to the gel. The closure may include a slot giving access to the medium to enable the zones to be cut out of the medium upon inspection of the image thereof on the film.

Ultraviolet imaging method and apparatus

An improved horizontal slab gel electrophoresis apparatus having compartments for vertical wicks at each end of a removable horizontal trough. Adjacent to the vertical wicks are vertical buffer compartments with built in electrodes. A partition separates the vertical wicks from the buffer compartment and includes a slot with a silicone plug which can be removed after curing or hardening of the vertical wicks to permit communication between the buffer and the gel slab. Flow adapters and an interconnecting tube permit constant circulation of the buffer to maintain equal volume and pH. The horizontal trough has slots for insertion of combs to form up to seventy-five sample wells and additional slots at each end for sealing the end of the horizontal slab when the gel slab is removed. The apparatus includes coils in a horizontal bed or plate beneath the trough for cooling the gel slab during a run and levelling screws for levelling the horizontal trough before pouring the gel slab.

Horizontal slab gel electrophoresis

An improved horizontal slab gel electrophoresis apparatus having compartments for vertical wicks at each end of a removable horizontal trough. Adjacent to the vertical wicks are vertical buffer compartments with built in electrodes. A partition separates the vertical wirks from the buffer compartment and includes a slot with a silicone plug which can be removed after curing or hardening of the vertical wicks to permit communication between the buffer and the gel slab. Flow adapters and an interconnecting tube permit constant circulation of the buffer to maintain equal volume and pH. The horizontal trough has slots for insertion of combs to form up to seventy-five sample wells and additional slots at each end for sealing the end of the horizontal slab when the gel slab is removed. The apparatus includes coils in a horizontal bed or plate beneath the trough for cooling the gel slab during a run and levelling screws for levelling the horizontal trough before pouring the gel slab.

Growing community of inventors

Malibu, CA, United States of America

Gary L Wilcox