Interleukin-1α antibodies and methods of use
Fully human monoclonal Abs includes (i) an antigen-binding variable region that exhibits very high binding affinity for IL-1α and (ii) a constant region that is effective at both activating the complement system though C1q binding and binding to several different Fc receptors.
1. A method for quantifying the amount of IL-1α in a biological sample, the method comprising the steps of:
(a) separating a biological sample obtained from a human subject using a size-exclusion filter that separates molecules according to molecular weight into a first fraction comprising intact IgG complexed with IL-1α and a second fraction consisting of molecules less than 100 Kda, wherein the biological sample comprises intact IgG complexed with IL-1α, wherein the first fraction does not pass through the filter, and wherein the second fraction passes through the filter; and
(b) quantifying the amount of IL-1α in the first fraction.
2. A method for quantifying the amount of IL-1α in a biological sample, the method comprising the steps of:
(a) separating a biological sample obtained from a human subject using a size-exclusion filter that separates molecules according to molecular weight into a first fraction comprising intact IgG complexed with IL-1α and a second fraction consisting of molecules less than 100 Kda, wherein the biological sample comprises intact IgG complexed with IL-1α, wherein the first fraction does not pass through the filter, and wherein the second fraction passes through the filter;
(b) adding the first fraction to a substrate comprising immobilized anti-human IgG antibodies (Abs) under conditions that allow IgG in the first fraction to specifically bind the anti-human IgG Abs immobilized on the substrate;
(c) washing the substrate to remove material in the first fraction that does not specifically bind the immobilized anti-human IgG Abs;
(d) contacting the substrate washed in step (c) with an antibody (Ab) that specifically binds human IL-1α under conditions that allows the Ab that specifically binds human IL-1α to specifically bind any human IL-1α bound to the substrate;
(e) washing the substrate to remove any of the Ab that specifically binds human IL-1α that is not bound to the substrate; and
(f) quantifying the amount of Ab that specifically binds human IL-1α remaining bound to the substrate after step (e).