This invention relates to compositions of matter, methods, modules and instruments for automated mammalian cell growth and mammalian cell transduction followed by nucleic acid-guided nuclease editing in live mammalian cells.

A fluidic system having a first volume, a second volume and a membrane geometrically separating the two volumes, which has an open-pore microstructure for the passage of a first medium and a second medium. There is a contact angle (Θ) between the interface of the media and the pore surface. A first electrical field in the region of the membrane and a first electromagnetic radiation and a first heating of the membrane define a first state (Z.sub.1), in which the membrane is not wetted or is less wetted by the first medium and is more heavily wetted by the second medium such that a first contact angle Θ.sub.1>90° is formed between the pore surface and the interface. The first medium and the second medium and the pore surface have a surface energy of which at least one surface energy can be reversibly changed in such a way that a second contact angle Θ.sub.2<Θ.sub.1 occurs between the pore surface and the interface in a second state (Z.sub.2).

The disclosure provides for compositions and methods of making and using a foam composition and its utility in clinical applications.

The invention provides a passive fluidics circuit for directing different fluids to a common volume, such as a reaction chamber or flow cell, without intermixing or cross contamination. The direction and rate of flow through junctions, nodes and passages of the fluidics circuit are controlled by the states of upstream valves (e.g. opened or closed), differential fluid pressures at circuit inlets or upstream reservoirs, flow path resistances, and the like. Free diffusion or leakage of fluids from unselected inlets into the common outlet or other inlets at junctions or nodes is prevented by the flow of the selected inlet fluid, a portion of which sweeps by the inlets of unselected fluids and exits the fluidics circuit by waste ports, thereby creating a barrier against undesired intermixing with the outlet flow through leakage or diffusion.

The disclosure relates to manufacturing a microfluidic arrangement wherein a second liquid (2) such as fluorinated oil is provided in direct contact with a continuous body of a first liquid (1) such as an aqueous cell culture medium and covering the first pot liquid. The second liquid is caused to move through the first liquid and into contact with a substrate (11) along all of a selected path to displace first liquid. The selected path is such that one or more walls of second liquid are formed that modify a shape of the continuous body of first liquid. The first liquid is aqueous. The second liquid is immiscible with the first liquid. The second liquid is treated in a liquid treatment apparatus (50), prior to the second liquid being caused to move through the first liquid, by flowing a gas through the second liquid and thereby increasing a level of saturation of the second liquid.

This invention relates to compositions of matter, methods, modules and instruments for automated mammalian cell growth and mammalian cell transduction followed by nucleic acid-guided nuclease editing in live mammalian cells.

A fluidic network for carrying out, in parallel, a plurality of analyses of biological samples is disclosed. The network has a flow cell array with a plurality of reaction chambers. The reaction chambers each have a first channel connection and a second channel connection. The first channel connections are connected to a first supply channel and the second channel connections are connected to a second supply channel. The first supply channel and the second channel connection are interconnected by a circulation line. At least one component is connected to the circulation line so that component test reagents can be introduced into the reaction chambers of the flow cell array.

Compositions of matter, methods, modules, and automated instruments may relate to synthesizing a library including an editing cassette including a different gRNA and donor DNA pair, amplifying the editing cassette in a partition separate from other editing cassettes in the library, adding nuclease to the partition, and adding lipofectamine to the editing cassette and nuclease to form a lipofectamine/nucleic acid/nuclease complex. A microcarrier coated in extracellular matrix or a cell adhesion molecule coating may be added to the lipofectamine/nucleic acid/nuclease complex. Cell growth material, the microcarrier, and mammalian cells may be transferred to a growth module in an automated closed cell editing instrument via a liquid handling system. The mammalian cells may be allowed to seed on the microcarrier. Conditions may be provided for the mammalian cells to take-up and be edited by a payload associated with the lipofectamine/nucleic acid/nuclease complex. The mammalian cells may be detached from the microcarrier.

Provided herein are compositions, systems, kits, and methods for detecting the presence or absence of a target protein in a discrete entity comprising: a) generating a discrete entity (e.g., microdroplet) comprising: i) a first cell that may secrete, or surface express, a target protein, ii) a quenched oligonucleotide probe, iii) first antibody-oligonucleotide conjugate or a particle-oligonucleotide conjugate, and a second antibody-oligonucleotide conjugate that bind the target protein in proximity to each to form an oligonucleotide template structure (OTS), and a nickase enzyme that cleaves the quenched oligonucleotide probe when it is hybridized to the OTS such that a detectable dye (e.g., fluorescent dye) is released and generates a signal; and b) detecting the presence or absence of the signal from the detectable dye.

The present invention provides an antibody including a structural domain represented by the following amino acid sequence, in an N- to C-direction, N-FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4-C
wherein the antibody further includes a protein molecule bound to the structural domain; the structural domain is capable of binding to a norovirus; FR denotes a framework region amino acid sequence and CDR denotes a complementary determining region amino acid sequence; any one of the following requirements (i)-(iii) is satisfied. Requirement (i): the CDR1 includes an amino acid sequence having a sequence identity of not less than 60% with any one of the amino acid sequences represented by SEQ ID NO: 1-SEQ ID NO: 6, the CDR2 includes an amino acid sequence having a sequence identity of not less than 60% with any one of the amino acid sequences represented by SEQ ID NO: 7-SEQ ID NO: 12, and the CDR3 includes an amino acid sequence having a sequence identity of not less than 60% with any one of the amino acid sequences represented by SEQ ID NO: 13-SEQ ID NO: 17; Requirement (ii): the CDR1 includes an amino acid sequence in which one-three amino acid(s) of any one of the amino acid sequence represented by SEQ ID NO: 1-SEQ ID NO: 6 has/have been substituted, deleted, or added, the CDR2 includes an amino acid sequence in which one-three amino acid(s) of any one of the amino acid sequence represented by SEQ ID NO: 7-SEQ ID NO: 12 has/have been substituted, deleted, or added, and the CDR3 includes an amino acid sequence in which one-three amino acid(s) of any one of the amino acid sequence represented by SEQ ID NO: 13-SEQ ID NO: 17 has/have been substituted, deleted, or added; and Requirement (iii): the CDR1 includes any one of the amino acid sequence represented by SEQ ID NO: 1-SEQ ID NO: 6, the CDR2 includes any one of the amino acid sequence represented by SEQ ID NO: 7-SEQ ID NO: 13, and the CDR3 includes any one of the amino acid sequence represented by SEQ ID NO: 13-SEQ ID NO: 17.