A neutralizing monoclonal antibody specific for the dimer interface region of IL-8.
We have generated two mAbs, 6G4.2.5 and A5.12.14, that are similarly capable of neutralizing the biologic activity of wild-type IL-8. To characterize these antibodies further, their reactivity against a series of engineered IL-8 monomer and dimer variants was examined using a neutrophil degranulation assay. While 6G4.2.5 was found to block effectively the biologic activity of all variants regardless of their dimerization status, the results for A5.12.14 differed dramatically. A5.12.14 fully inhibited the agonist activity of one of the monomer variants, partially blocked the activity of another, and had no effect on the activity of two other variants. These results suggested that the binding epitope of A5.12.14 was being affected by the particular amino acid substitutions introduced into the dimer interface region of the variants to disfavor dimerization. If A5.12.14 indeed binds to the dimer interface region of IL-8, it could be predicted that this mAb would be unable to inhibit the activity of dimeric IL-8. This was confirmed in studies which showed that A5.12.14 had no demonstrable effect on the activity of a constitutively dimeric IL-8 variant. These studies represent the first example of a mAb specific for the dimerization status of IL-8.
Deforge
L E
LE
Departments of BioAnalytical Technology, Genentech, Inc., South San Francisco, CA 94080, USA. deforge.laura@gene.com
Lowman
H B
HB
Leong
S R
SR
Chuntharapai
A
A
Jin Kim
K
K
Hébert
C A
CA
D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D000911:Antibodies, Monoclonal; D002118:Calcium / Q000378:metabolism; D019281:Dimerization; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D005966:Glucuronidase / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism*; D051379:Mice; D009504:Neutrophils / Q000201:enzymology; D011485:Protein Binding
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000939:Epitopes; D007074:Immunoglobulin G; D016209:Interleukin-8; D005966:Glucuronidase; D002118:Calcium