The invention pertains to a single polypeptide chain binding molecule which has binding specificity and affinity substantially similar to the binding specificity and affinity of the light and heavy chain aggregate variable region of an antibody, to genetic sequences coding therefor, and to recombinant DNA methods of producing such molecule and uses for such molecule.

Single polypeptide chain binding molecules

A member of a specific binding pair (sbp) is identified by expressing DNA encoding a genetically diverse population of such sbp members in recombinant host cells in which the sbp members are displayed in functional form at the surface of a secreted recombinant genetic display package (rgdp) containing DNA encoding the sbp member or a polypeptide component thereof, by virtue of the sbp member or a polypeptide component thereof being expressed as a fusion with a capsid component of the rgdp. The displayed sbps may be selected by affinity with a complementary sbp member, and the DNA recovered from selected rgdps for expression of the selected sbp members. Antibody sbp members may be thus obtained, with the different chains thereof expressed, one fused to the capsid component and the other in free form for association with the fusion partner polypeptide. A phagemid may be used as an expression vector, with said capsid fusion helping to package the phagemid DNA. Using this method libraries of DNA encoding respective chains of such multimeric sbp members may be combined, thereby obtaining a much greater genetic diversity in the sbp members than could easily be obtained by conventional methods.

Methods for producing members of specific binding pairs

An altered antibody is produced by replacing the complementarity determining regions (CDRs) of a variable region of an immunoglobulin (Ig) with the CDRs from an Ig of different specificity, using recombinant DNA techniques. The gene coding sequences for producing the altered antibody may be produced by site-directed mutagenesis using long oligonucleotides.

Recombinant altered antibodies and methods of making altered antibodies

The present invention concerns polypeptides comprising a variant Fc region. More particularly, the present invention concerns Fc region-containing polypeptides that have altered effector function as a consequence of one or more amino acid modifications in the Fc region thereof.

Polypeptide variants with altered effector function

Multichain polypeptides or proteins and processes for their production in cells of host organisms which have been transformed by recombinant DNA techniques. According to a first aspect of the present invention, there is provided a process for producing a heterologous multichain polypeptide or protein in a single host cell, which comprises transforming the host cell with DNA sequences coding for each of the polypeptide chains and expressing said polypeptide chains in said transformed host cell. According to another aspect of the present invention there is provided as a product of recombinant DNA technology an Ig heavy or light chain or fragment thereof having an intact variable domain. The invention also provides a process for increasing the level of protein expression in a transformed host cell and vectors and transformed host cells for use in the processes.

Multichain polypeptides or proteins and processes for their production

Altered and native immunoglobulins, including constant-variable region chimeras, are prepared in recombinant cell culture. The immunoglobulins contain variable regions which are immunologically capable of binding predetermined antigens. Methods are provided for refolding directly expressed immunoglobulins into immunologically active form.

Recombinant immunoglobin preparations

The invention relates to processes for producing an immunoglobulin or an immunologically functional immunoglobulin fragment containing at least the variable domains of the immunoglobulin heavy and light chains. The processes can use one or more vectors which produce both the heavy and light chains or fragments thereof in a single cell. The invention also relates to the vectors used to produce the immunoglobulin or fragment, and to cells transformed with the vectors.

Methods of producing immunoglobulins, vectors and transformed host cells for use therein

PURPOSE: To obtain a recombinant immunoglobulin having improved cell treatment characteristics and antigen-binding properties by transforming a host with an expression vector into which a heavy chain or a light chain of immunoglobulin modified in a predetermined manner is inserted and culturing the host. CONSTITUTION: A DNA molecule comprising a sequence coding for a heavy chain or a light chain of a changed immunoglobulin having specificity to a prescribed identified antigen, which is modified in a predetermined manner by an alteration, a deletion or an addition of at least one amino acid sequence from an amino acid sequence of mammalian immunoglobulin is prepared. At least one host cell is transformed with at least one replicable expression vector containing the sequence operatively bonded to a proper promoter, an immunoglobulin is recovered by the culture of the host cell to give the objective changed immunoglobulin containing a constant and a variable regions, having specificity to a specified identified antigen. COPYRIGHT: (C)1995,JPO

Shmuel Cabilly

Papers

Recombinant immunoglobin preparations

Methods of producing immunoglobulins, vectors and transformed host cells for use therein

Recombinant immunoglobulin preparations

Methods of making antibody heavy and light chains having specificity for a desired antigen

Altered recombinant immunoglobulins