US 11,933,778 B2
Lipid-free anchoring of thermophilic bacteriophage G20C portal adapter into solid-state nanopores
Meni Wanunu, Needham, MA (US); Alfred Antson, York (GB); Sandra Greive, York (GB); and Benjamin Cressiot, Montlignon (FR)
Assigned to Northeastern University, Boston, MA (US); and University of York, York (GB)
Filed by Northeastern University, Boston, MA (US); and University of York, York (GB)
Filed on Jun. 29, 2022, as Appl. No. 17/809,705.
Application 17/809,705 is a continuation of application No. 16/416,139, filed on May 17, 2019, granted, now 11,408,880.
Claims priority of provisional application 62/673,118, filed on May 17, 2018.
Prior Publication US 2022/0373533 A1, Nov. 24, 2022
Int. Cl. G01N 33/487 (2006.01); B81B 1/00 (2006.01); B82Y 15/00 (2011.01); C07K 14/005 (2006.01); C12Q 1/6869 (2018.01)
CPC G01N 33/48721 (2013.01) [B81B 1/002 (2013.01); C07K 14/005 (2013.01); C12Q 1/6869 (2013.01); B82Y 15/00 (2013.01); C12N 2795/00022 (2013.01); Y10S 930/22 (2013.01)] 15 Claims
OG exemplary drawing
 
1. A protein variant, the protein variant comprising a modification of an amino acid sequence comprising SEQ ID NO: 1, the modification comprising one or more of:
(i) a modification of an upper internal surface residue or a lower internal surface residue of the protein variant, the modification comprising an alteration of an electrostatic surface potential of an upper internal surface or a lower internal surface of SEQ ID NO: 1;
(ii) a modification comprising an insertion of a cysteine residue into SEQ ID NO: 1 or a replacement of an amino acid residue of SEQ ID NO: 1 with a cysteine residue;
(iii) a modification of a tunnel loop residue of the protein variant, the modification comprising an expansion of a narrowest constriction of a tunnel loop of SEQ ID NO: 1, a restriction of the narrowest constriction of the tunnel loop of SEQ ID NO: 1, or a removal of the narrowest constriction of the tunnel loop of SEQ ID NO: 1;
(iv) a modification of a tunnel loop residue of the protein variant, the modification resulting in an alteration of an electrostatic charge property of a tunnel loop of SEQ ID NO: 1;
(v) a modification which alters an external charge of the protein variant;
(vi) a modification which promotes binding of the protein variant to a solid-state matrix;
(vii) a modification which extends an N-terminus or a C-terminus of SEQ ID NO: 1 or an N-terminus or a C-terminus of a cleaved portion of SEQ ID NO: 1; and
(viii) a deletion of an amino acid residue of at least one of a C-terminal region and an N-terminal region of SEQ ID NO: 1.