In vitro evolution in microfluidic systems
View Patent ↗The invention describes a method for isolating one or more genetic elements encoding a gene product having a desired activity, comprising the steps of: (a) compartmentalizing genetic elements into microcapsules; and (b) sorting the genetic elements which express the gene product having the desired activity; wherein at least one step is under microfluidic control. The invention enables the in vitro evolution of nucleic acids and proteins by repeated mutagenesis and iterative applications of the method of the invention.
1. A method for detecting a product of an enzymatic reaction, comprising the steps of:
providing a droplet generator to produce, under microfluidic control, a plurality of aqueous microcapsules surrounded by an immiscible continuous phase that comprises a fluorinated oil that comprises a fluorinated polymer surfactant, each of the plurality of microcapsules comprising an enzyme, a genetic element, and reagents for the enzymatic reaction;
pooling the microcapsules into one or more common compartments such that a portion of the plurality of microcapsules contact each other but do not fuse with each other due to the presence of the surfactant;
conducting the enzymatic reaction on the genetic element of at least one of the plurality of microcapsules within the one or more common compartments; and
detecting the product of the enzymatic reaction.
2. The method of claim 1 , wherein the genetic elements are nucleic acids, proteins, or cells.
3. The method of claim 2 , wherein the nucleic acids are primers for a polymerase chain reaction (PCR).
4. The method of claim 1 , wherein the genetic elements are labeled.
5. The method of claim 4 , wherein the genetic elements are optically labeled.
6. The method of claim 4 , wherein the genetic elements are fluorescently labeled antibodies.
7. The method of claim 4 , wherein the genetic elements are chemically labeled antibodies.
8. The method of claim 1 , wherein the microcapsules are monodisperse with respect to each other.
9. The method of claim 1 , wherein the droplet generator further comprises an aqueous fluid channel and two immiscible continuous phase channels and producing the plurality of microcapsules surrounded by the immiscible continuous phase under the microfluidic control comprises partitioning an aqueous fluid that is flowing through the aqueous fluid channel with two counter propagating streams of the immiscible continuous phase that are flowing through the immiscible continuous phase channels.