US 5,858,728 A · Monoclonal antibody against LPS core

Abstract

The invention provides monoclonal antibodies (Mabs) which are cross-protective against endotoxemia caused by at least two different Gram-negative bacterial strains having different core structures; and methods of production of these antibodies. By use of the Kohler/Milstein procedure involving immunization of mice with a number of different rough strains of heat-killed Gram-negative bacteria, followed by fusion and proper screening of the resulting hybridomas, such murine MAbs are obtained. The murine MAbs may be chimerized or humanized by known methods. For example, a chimeric MAb of IgG isotype is provided in which the hypervariable regions of the heavy chain have the amino acid sequences: Asp Tyr Tyr Met Thr; Leu Ile Arg Asn Lys Arg Asn Gly Asp Thr Ala Glu Tyr Ser Ala Ser Val Lys; and Gln Gly Arg Gly Tyr Thr Leu Asp Tyr; the hypervariable regions of the light chain have the amino acid sequences: Arg Ala Ser Gln Asn Ile Asn Ile Trp Leu Ser; Lys Ala Ser Asn Leu His Thr; and Leu Gln Gly Gln Ser Tyr Pro Arg Thr; the framework regions in the variable domains are murine and the constant domains are human.

Claims26

Dependent claims

A monoclonal antibody which binds an epitope in the core region of the LPS molecule and which is cross-reactive to, and cross-protective against endotoxemia caused by, at least two different Gram-negative bacterial strains having different core structures, said bacterial strains selected from the group consisting of:

(a) smooth E. coli,
(b) rough mutant E. coli of core types R1, R2, R3, R4 and K12;
and
(c) Salmonella;
said monoclonal antibody having a constant region of human origin, a variable framework region of human or non-human origin, and a hypervariable region of non-human origin.

2 more elements

An LPS binding molecule which comprises at least one antigen binding site comprising at least one domain which comprises in sequence framework region hFR1, hypervariable region hCDR1, framework region hFR2, hypervariable region hCDR2, framework region hFR3, hypervariable region hCDR3, and framework region hFR4;

said hCDR1 having the amino acid sequence Asp Tyr Tyr Met Thr which are amino acids 31-35 of SEQ ID NO:2 or SEQ ID NO:4;
said hCDR2 having the amino acid sequence 50-67 of SEQ ID NO:2
wherein amino acid 54 is Lys or Tyr and amino acid 67 is Lys or Arg;
said hCDR3 having the amino acid sequence Gln Gly Arg Gly Tyr Thr Leu Asp Tyr which are amino acids 101-109 of SEQ ID NO:2 or SEO ID NO:4;
and direct equivalents thereof, said LPS binding molecule which binds an epitope in the core region of the LPS molecule of gram negative bacterial strains having different core structures and selected from the group consisting of smooth E. coli;
rough E. coli of core types R1, R2, R3, R4 and K12;
and Salmonella.

4 more elements

Dependencies are inferred from the claim text.

Monoclonal antibody against LPS core

Life of this patent

Monoclonal antibody against LPS core

Life of this patent

Abstract

Claims26