Techniques for predicting, detecting and reducing aspecific protein interference in assays involving immunoglobulin single variable domains

Abstract

This invention provides, and in certain specific but non-limiting aspects relates to: assays that can be used to predict whether a given ISV will be subject to protein interference as described herein and/or give rise to an (aspecific) signal in such an assay (such as for example in an ADA immunoassay). Such predictive assays could for example be used to test whether a given ISV could have a tendency to give rise to such protein interference and/or such a signal; to select ISV's that are not or less prone to such protein interference or to giving such a signal; as an assay or test that can be used to test whether certain modification(s) to an ISV will (fully or partially) reduce its tendency to give rise to such interference or such a signal; and/or as an assay or test that can be used to guide modification or improvement of an ISV so as to reduce its tendency to give rise to such protein interference or signal; —methods for modifying and/or improving ISV's to as to remove or reduce their tendency to give rise to such protein interference or such a signal; —modifications that can be introduced into an ISV that remove or reduce its tendency to give rise to such protein interference or such a signal; ISV's that have been specifically selected (for example, using the assay(s) described herein) to have no or low(er)/reduced tendency to give rise to such protein interference or such a signal; modified and/or improved ISV's that have no or a low(er)/reduced tendency to give rise to such protein interference or such a signal.

1 . (canceled) 
     
     
         2 . A monovalent heavy chain immunoglobulin single variable domain (ISVD) that terminates in the sequence VTVSS(X) n  (SEQ ID NO: 34), in which n is 1 and X is alanine (A). 
     
     
         3 . The monovalent heavy chain ISVD of  claim 2 , wherein the monovalent heavy chain ISVD does not comprise a valine (V) a position 11. 
     
     
         4 . The monovalent heavy chain ISVD of  claim 2 , comprising a leucine (L) at Kabat position 11. 
     
     
         5 . The monovalent heavy chain ISVD of  claim 2 , comprising a serine (S) at Kabat position 11. 
     
     
         6 . The monovalent heavy chain ISVD of  claim 2 , wherein the monovalent heavy chain ISVD does not comprise a proline (P) at Kabat position 14. 
     
     
         7 . The monovalent heavy chain ISVD of  claim 2 , comprising an alanine (A) at Kabat position 14. 
     
     
         8 . The monovalent heavy chain ISVD of  claim 2 , wherein the monovalent heavy chain ISVD does not comprise an arginine (R) at Kabat position 83. 
     
     
         9 . The monovalent heavy chain ISVD of  claim 2 , comprising a lysine (K) at Kabat position 83. 
     
     
         10 . The monovalent heavy chain ISVD of  claim 2 , wherein the monovalent heavy chain ISVD does not comprise a lysine at Kabat position 108. 
     
     
         11 . The monovalent heavy chain ISVD of  claim 2 , comprising a glutamine (Q) at Kabat position 108. 
     
     
         12 . The monovalent heavy chain ISVD of  claim 2 , wherein the monovalent heavy chain ISVD does not bind serum albumin. 
     
     
         13 . The monovalent heavy chain ISVD of  claim 2 , wherein the monovalent heavy chain ISVD binds a therapeutically relevant target. 
     
     
         14 . The monovalent heavy chain ISVD of  claim 2 , wherein the monovalent heavy chain ISVD is a VHH domain. 
     
     
         15 . The monovalent heavy chain ISVD of  claim 14 , wherein the VHH domain is a humanized or sequenced optimized VHH domain. 
     
     
         16 . The monovalent heavy chain ISVD of  claim 2 , wherein the monovalent heavy chain ISVD is a VH domain. 
     
     
         17 . The monovalent heavy chain ISVD of  claim 16 , wherein the VH domain is a human VH domain or a camelized VH domain. 
     
     
         18 . A pharmaceutical composition comprising the monovalent heavy chain ISVD of  claim 2 , and at least one carrier, diluent or excipient. 
     
     
         19 . A polypeptide comprising the monovalent heavy chain IS VD of  claim 2  at its C-terminal end, wherein the polypeptide is a not a multivalent polypeptide. 
     
     
         20 . A pharmaceutical composition comprising the polypeptide of  claim 19 , and at least one carrier, diluent or excipient.