Method of isolating human antibodies
AssigneeImmunoQure AG
InventorsAdrian Hayday, Kai Krohn, Annamari Ranki, Part Peterson, Kai Kisand, Edward Stuart, Annalisa Macagno
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Abstract
Provided is a novel method of isolating and producing human antibodies with desired specificity from human B cells. In particular, a method of isolating human antibodies from memory B cells obtained from patients which suffer from a disease which is caused by or involves activation of the immune system, for example autoimmune and inflammatory disorders is described.
Claims14
A method of producing a human antibody or binding fragment thereof with desired specificity characterized by isolating B cells from short term oligoclonal cultures of activated B cells that secrete antibodies of IgG isotype comprising:
- (a) selecting memory B cells that express antibodies against a protein of interest from one or more biological samples derived from a subject suffering from an autoimmune disease, whose sera is screened for the presence of auto-antibodies against a protein of interest, on the basis of the expression of at least one of CD22 cell-surface membrane marker and antigen binding;
- (b) stimulating the selected cells for about three to five hours with a first polyclonal B cell activator which is Epstein-Barr virus (EBV) under cell culture conditions and in the absence of a second polyclonal B cell activator;
- (c) separating the cells from said activator;
- (d) activating the stimulated cells with a second polyclonal B cell activator which is a CpG-based oligonucleotide under cell culture conditions;
- and,
- (e) screening the activated cells to identify cells that express IgG isotype antibodies of interest, wherein the culture conditions of step (d) do not comprise a cytokine.
Dependencies are inferred from the claim text.