Berkeley Lights, Inc.

Bruker Cellular Analysis, Inc.

Jason M McEwen

Randall D Lowe, Jr

Mark P White

Kevin T Chapman

Gang F Wang

J Tanner Nevill

Volker L S Kurz

Alexander J Mastroianni

Xiao Guan Radstrom

Xiao Guan

Daniele Malleo

Andrew W McFarland

Aathavan Karunakaran

Xiaohua Wang

Kristin G Beaumont

Natalie C Marks

Kellen C Mobilia

Keith J Breinlinger

Jian Gong

Justin K Valley

Gregory G Lavieu

Minha Park

Peggy A Radel

Guido K Stadler

Shao Ning Pei

Annamaria Mocciaro

Magali Soumillon

Patricia A Dyck

Ravi K Ramenani

George L Fox

Joshua J Cardiel Rivera

Troy A Lionberger

Keith Joseph Breinlinger

Eric K Sackmann

X Robert Bao

Brian A Rabkin

Christine E Sun

Systems, methods, and kits therefor, enabling rapid protein evolution are described herein. A system useful in the methods described herein include a DNA synthesis component; a microfluidic system including a microfluidic device having a microfluidic channel and sequestration pens; and a computing component, which is configured to analyze assay results and, based upon the analysis, design improved DNA sequences for iterative protein evolution. The microfluidic system is configured to permit correlation of DNA sequence on a bead to its location within the microfluidic device, permit cell free protein expression of a DNA sequence captured to a bead, and to permit assay of the protein so produced.

Systems and methods for identification of optimized protein production and kits therefor

In biosciences and related fields, it can be useful to study cells in isolation so that cells having unique and desirable properties can be identified within a heterogenous mixture of cells. Processes and methods disclosed herein provide for encapsulating cells within a microfluidic device and assaying the encapsulated cells. Encapsulation can, among other benefits, facilitate analyses of cells that generate secretions of interest which would otherwise rapidly diffuse away or mix with the secretions of other cells.

Methods for encapsulating and assaying cells

Systems, methods and kits are described for culturing one or more biological cells in a microfluidic device, including provision of nutrients and gaseous components configured to enhance cell growth, viability, portability, or any combination thereof. In some embodiments, culturing a single cell may produce a clonal population in the microfluidic device.

Microfluidic cell culture

Reagents for stabilizing the nucleic acids of a biological cell, compositions, kits and methods of use thereof are described. The stabilization reagents may prepare the nucleic acids within the biological cell for storage and preserve the representative population of the nucleic acids for later isolation and analysis.

Nucleic acid stabilization reagent, kits, and methods of use thereof

A method of preparing an antibody therapeutic is provided comprising: (a) providing a dissociated cell sample from at least one solid tumor sample obtained from a patient; (b) loading the dissociated cell sample into a microfluidic device having a flow region and at least one isolation region fluidically connected to the flow region; (c) moving at least one B cell from the dissociated cell sample into at least one isolation region in the microfluidic device, thereby obtaining at least one isolated B cell; and (d) using the microfluidic device to identify at least one B cell that produces antibodies capable of binding to cancer cells. The cancer cells can be the patient's own cancer cells. Also provided are methods of treating patients, methods of labeling or detecting cancer, engineered T or NK cells comprising antibodies or fragments thereof, and engineered antibody constructs.

Methods of producing patient-specific anti-cancer therapeutics and methods of treatment therefor

Microfluidic devices having an electrowetting configuration and an optimized droplet actuation surface are provided. The devices include a conductive substrate having a dielectric layer, a hydrophobic layer covalently bonded to the dielectric layer, and a first electrode electrically coupled to the dielectric layer and configured to be connected to a voltage source. The microfluidic devices also include a second electrode configured to be connected to the voltage source. The hydrophobic layer features self-associating molecules covalently bonded to a surface of the dielectric layer in a manner that produces a densely-packed monolayer that resists intercalation and or penetration by polar molecules or species. Also provided are microfluidic devices having an electrowetting configuration that further include a section having a dielectrophoresis configuration; systems that include microfluidic devices in combination with an aqueous droplet and a fluidic medium immiscible with the aqueous droplet; related kits; and methods of manipulating droplets within the microfluidic devices.

Microfluidic apparatus having an optimized electrowetting surface and related systems and methods

Methods are described herein for isolating clonal populations of cells having a defined genetic modification. The methods are performed, at least in part, in a microfluidic device comprising one or more sequestration pens. The methods include the steps of: maintaining individual cells (or precursors thereof) that have undergone a genomic editing process in corresponding sequestration pens of a microfluidic device; expanding the individual cells into respective clonal populations of cells; and detecting, in one or more cells of each clonal population, the presence of a first nucleic acid sequence that is indicative of the presence of an on-target genome edit in the clonal population of cells. Also described are methods of performing genome editing within a microfluidic device, and compositions comprising one or more clonal populations of cells generated according to the methods disclosed herein.

Methods, systems and devices for selection and generation of genome edited clones

Microfluidic devices having an electrowetting configuration and an optimized droplet actuation surface are provided. The devices include a conductive substrate having a dielectric layer, a hydrophobic layer covalently bonded to the dielectric layer, and a first electrode electrically coupled to the dielectric layer and configured to be connected to a voltage source. The microfluidic devices also include a second electrode, optionally included in a cover, configured to be connected to the voltage source. The hydrophobic layer features self-associating molecules covalently bonded to a surface of the dielectric layer in a manner that produces a densely-packed monolayer that resists intercalation and or penetration by polar molecules or species. Also provided are microfluidic devices having an electrowetting configuration that further include a section or module having a dielectrophoresis configuration; systems that include any of the microfluidic devices in combination with an aqueous droplet and a fluidic medium immiscible with the medium of the aqueous droplet; related kits; and methods of manipulating droplets, optionally containing micro-objects such as biological cells, within the microfluidic devices.

Systems and methods are described herein for improved droplet generation within microfluidic apparatuses. Electrowetting forces of varying configurations may be used to separate droplets from a fluidic reservoir in a reproducible and rapid manner. In many embodiments, separation of droplets from the fluidic reservoir is performed without the use of highly specialized surfactants.

Droplet generation in a microfluidic device having an optoelectrowetting configuration

Functional assays using reporter cell assays are described which probe the activity of at least one cell of interest. The ability to probe at least one cell is provided by using the microfluidic methods, devices and kits described herein. Assays combining both reporter cell signaling as well as binding assay signaling for at least one cell is also described herein.

Microfluidic reporter cell assay methods and kits thereof

Growing community of inventors

El Cerrito, CA, United States of America

Jason M McEwen