Methods and compositions for assembly of biological nanopores

A method of making a detection apparatus comprising one or more native nanopore proteins, comprising the steps of:

• (a) forming an aqueous mixture comprising a nanopore protein, a membrane scaffold protein (MSP), and a first lipid to produce a sample of nanodisc-nanopore protein complexes, wherein a population of the nanodisc-nanopore protein complexes in the sample each comprise a native nanopore protein;
• (b) providing a solid support comprising one or more apertures, wherein a membrane is formed over each of the apertures, wherein the membrane comprises a second lipid, and wherein the membrane separates a cis chamber from a trans chamber in the detection apparatus;
• and
• (c) contacting the one or more membranes with the population of nanopore-nanodisc complexes comprising the native nanopore protein to assimilate a native nanopore protein into each of the membranes;
• wherein the aqueous mixture further comprises a detergent, wherein the final concentration of the detergent is from about 14 mM to about 40 mM;
• wherein the first lipid is 1.2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC), the MSP is MSP1D1, or a variant thereof, the nanopore protein is α-hemolysin (α-HL) or a variant thereof, the detergent is cholate, and the second lipid is 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (DPhPE);
• and
• wherein the molar ratio of first lipid to MSP to nanopore protein is about 101:6:1 or about 120:6:1.