Harvard College

Ultivue, Inc.

Rootpath Genomics, Inc.

Guangzhou Chengyuan Bioimmunology Technology Co., Ltd.

Xi Chen

Peng Yin

Ely Blanton Porter

Stephanie Rae Hennek

Ralf Jungmann

Sarit Agasti

Michael J Natan

Mael Manesse

Mingjie Dai

Feng Xuan

Abdul Majeed Mohammed

David Y Zhang

Thomas E Schaus

David Yu Zhang

The present disclosure describes various improved methods for imaging at least one target in a sample, including methods employing an adapter strand oligonucleotide and a bridge strand oligonucleotide. Some methods also employ bouncer oligonucleotides and/or blocker oligonucleotides. Some methods also use two partial docking strands to detect proximity of the partial docking strands to each other.

Multiplexed imaging using strand displacement

Provided herein are compositions and methods for assembling nucleic acid sequences encoding T-cell receptors.

Compositions and methods for t-cell receptor gene assembly

The present application provides certain advantageous ways of conducting multiplexed imaging.

Methods for multiplex imaging using labeled nucleic acid imaging agents

The present disclosure provides, in some embodiments, methods and compositions for single-molecule detection.

Conditional primer extension for single-molecule detection

Provided herein are compositions and methods to identify a binding element (e.g., peptide, peptoid, or protein) that can be bound by an immunoreceptor (e.g., antibody). The binding element can be provided in a target binding unit comprising two binding elements separated by a spacer such that the two binding elements simultaneously bind to a single molecule comprising an antigen binding domain of an antibody. The present disclosure provides various methods to construct the spacer. The identified binding elements can be further used to manufacture an array which can be used to profile antibodies obtained from a blood sample.

Target binding moiety compositions and methods of use

Provided herein are compositions and methods for high throughout cloning of fused bipartite immunoreceptor polynucleotides encoding cognate pairs of bipartite immunoreceptors. Also provided herein are various applications of the fused bipartite immunoreceptor polynucleotides, expression vectors containing the fused bipartite immunoreceptor polynucleotides, or cells containing the fused bipartite immunoreceptor polynucleotides or expression vectors.

High throughput cloning of paired bipartite immunoreceptor polynucleotides and applications thereof

Compositions and methods for T-cell receptor gene assembly

Provided herein, in some aspects, are methods of imaging molecules without a microscope or other specialized equipment, referred to herein as 'microscope-free imaging (MFI).' Herein, 'molecular instruments' (e.g., DNA-based and protein-based molecules) are used, instead of microscopes, in a &#x201c;bottom-up&#x201d; approach for inspecting molecular targets.

Microscope-free imaging

A method for exchange imaging of at least two targets in a sample includes (a) incubating a sample with at least two or more target-recognizing antibodies, each bound to a corresponding monovalent tight antibody binder-docketing moiety (MTAB-DM) reagent capable of binding monovalently to the target-recognizing antibodies, (b) applying at least two imager moieties corresponding to the MTAB-DM, either in series, in batches, or in parallel, and (d) imaging the at least two imager moieties either in series, in batches, or in parallel.

Compositions and methods to expedite antibody-based exchange imaging

An imaging agent comprises (a) at least one target recognition moiety; (b) at least one observable moiety non-transiently bound to the target recognition moiety, and (c) at least one docking moiety bound to the target recognition moiety, wherein the docking moiety is capable of transiently binding at least one observable moiety. In some embodiments, the at least one target recognition moiety is an antibody or antigen binding fragment thereof. A method of performing super-resolution imaging on a sample comprises (a) contacting the sample with at least one imaging agent comprising (i) a target-recognizing molecule non-transiently attached to at least one observable moiety and (ii) a target-recognizing molecule attached to at least one docking moiety, wherein the docking moiety is capable of transiently binding at least one observable moiety; (b) imaging the target-recognizing molecule non-transiently bound to at least one observable moiety; and (c) providing at least one observable moiety capable of transiently binding to the docking moiety and imaging the observable moiety transiently bound to the docking moiety.

Growing community of inventors

West Newton, MA, United States of America

Xi Chen