Removal of amphipathic epitopes from genetically engineered antibodies: production of modified immunoglobulins with reduced immunogenicity.
Several approaches have been developed to reduce the human immune response to nonhuman antibodies. However, chimeric antibodies and humanized antibodies often have decreased binding affinity. We described a new approach for reducing the immunogenicity of chimeric antibodies while maintaining the affinity. This approach seeks to prevent the recognition of murine immunogenic peptides from the antibody variable region by human lymphocytes. Putative immunogenic epitopes in the variable region are identified and subjected to site directed mutagenesis to make them human and/or to break the amphipathic motifs. The R3 antibody, which blocks the epidermal growth factor (EGF) receptor, was used as a model system to test this approach. Four segments containing possible amphipathic epitopes were found in the heavy variable domain using the program AMPHI. Six amino acids within two of these segments were substituted by the corresponding residues from a homologous human sequence. No mutations were made in the murine light variable domain. Experiments in monkeys suggested that the "detope" R3 antibody was less immunogenic than its chimeric analogue. A search for possible amphipathic epitopes in the Kabat database revealed the presence of conserved patterns in the different families of variable region sequences, suggesting that the proposed method may be of general applicability.
Mateo
C
C
Center of Molecular Immunology, Havana, Cuba. cristina@ict.cim.sld.cu
Lombardero
J
J
Moreno
E
E
Morales
A
A
Bombino
G
G
Coloma
J
J
Wims
L
L
Morrison
S L
SL
Pérez
R
R
D000465:Algorithms; D000818:Animals; D000906:Antibodies / Q000235:genetics* / Q000276:immunology / Q000494:pharmacology*; D000918:Antibody Specificity; D002522:Chlorocebus aethiops; D022801:Complementarity Determining Regions / Q000276:immunology; D000939:Epitopes / Q000737:chemistry / Q000276:immunology*; D066246:ErbB Receptors / Q000276:immunology; D006801:Humans; D007114:Immunization; D051379:Mice; D008969:Molecular Sequence Data; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid
D000906:Antibodies; D022801:Complementarity Determining Regions; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D066246:ErbB Receptors