Pierce Biotechnology, Inc.

Thermo Finnigan LLC

Christopher Laurence Etienne

Kay K Opperman

Barbara J Kaboord

Jean-Samuel Schultz

Scott D Meier

Ramesh Ganapathy

Paul Jeffrey Haney

Erum Raja

Sijian Hou

Michael W Senko

Greg Hermanson

Graeme C McAlister

John Charles Rogers

Eric Leigh Hommema

Ryan Daniel Bomgarden

Ashok Salunkhe

Atul Deshpande

Deven Etnyre

Graeme Mcalister

Jeffrey Paul Haney

The present specification relates to compositions, devices, apparatus, methods, kits and systems for sample preparation (e.g., separation, reduction or removal of small molecules from biomolecules in a sample). Exemplary small molecules that can be separated, reduced or removed have a molecular weight range of <2000 Da. and may include, but are not limited to, dyes, biotin, affinity tags, crosslinkers, reducing agents, labels, nanoparticles, radioactive ligands, mass tags, unreacted molecules and combinations, intermediates and derivatives of the foregoing. Exemplary biomolecules present in a sample, include but are not limited to, proteins, glycoproteins, antibodies, peptides, nucleic acids, polysaccharides, carbohydrates and lipids. Methods, compositions, kits, devices, apparatus and systems of the disclosure may advantageously provide superior separation of small molecule contaminants and additionally reduce time and expenses related to separation of small molecules from larger biomolecules in samples. Biomolecules separated as set forth herein are amenable to better downstream processing.

Sample preparation compositions, devices, systems and methods

Peptide and/or protein quantitation methods, kits, and compositions, particularly useful for mass spectrometry, are provided herein based on a bathocuproine-based composition complex such as bathocuproinedisulfonic acid disodium salt hydrate complex. The methods are one-step rapid absorbance methods using small sample volumes. They produce a robust signal with high signal to background ratio and accurately quantitate even complex peptide mixtures with low variability and high sensitivity.

Quantitative peptide or protein assay

Isobaric mass spectrometry tags (e.g., TMT) are susceptible to ratio compression, which arises from the co-isolation and co-fragmentation of interfering species that also contribute to the final reporter ion ratios. Additional stages of ion activation/transformation (e.g., MSn and PTR) have been shown to decrease ratio compression. Embodiments of the present invention include a mass spectrometry cleavable moiety on the isobaric mass tags. The cleavable moiety allows for a predictable mass loss, and results in an improved tag reporter ion purity.

Methods of mass spectrometry quantitation using cleavable isobaric tags and neutral loss fragmentation

Modified nucleotides, and methods to modify nucleotides with a moiety or label, such as biotin, that permits their detection and results in a modified nucleotide, and methods of use of the modified nucleotide in quantitative and qualitative assays.

Modified nucleotides methods and kits

Methods and sets of non-biological reagents (elution reagents, tag isomers, tag reactive groups, crosslinkers) for single or multiplexed capture and gentle elution of biomolecules. Examples are provided using amine- and cysteine-reactive reagents for enrichment of proteins, peptides, and rare peptide modifications.

Methods and reagents for biomolecule labeling, enrichment and gentle elution

Modified nucleotides

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Fitchburg, WI, United States of America

Christopher Laurence Etienne