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Ensuring the Safety and Efficacy of RNA Therapeutics Through double stranded RNA (dsRNA) Impurity Control

A critical challenge faced during the development and manufacturing of RNA-based therapies is the presence of unintended impurities, particularly double-stranded RNA (dsRNA). While dsRNA is an inevitable byproduct of the in vitro transcription (IVT) process, unchecked contamination can severely jeopardize both the safety and effectiveness of the treatment

5 days ago
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Protocol for Conjugation of a Biomolecule to Aldehyde Particles

This protocol describes a method for coupling ligands containing hydrazine, hydrazide, aminooxy, or amine groups to aldehyde-containing particles. Aldehyde particles are reactive with hydrazine, hydrazide, or aminooxy derivatives, forming hydrazone or oxime linkages. Reactions with amine-containing molecules, such as proteins, can be carried out through a reductive amination process using sodium cyanoborohydride.

8 months ago
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Protocol for Conjugation of a Biomolecule to Epoxy Particles

This protocol describes a method for coupling ligands containing thiol, amino, or hydroxyl groups to epoxide-containing particles via a ring-opening reaction that is facilitated under alkaline conditions. This reactive group can be used to couple proteins, nucleic acids, sugars and carbohydrates, and other organic molecules containing these functionalities.

8 months ago
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Protocol for Conjugation of a Biomolecule to Hydroxyl Particles

This protocol describes methods for covalently binding ligands to hydroxyl particles. Since hydroxyl groups are not spontaneously reactive with biomolecules, they must be activated using a variety of methods. Most activation strategies are carried out under nonaqueous conditions because the activating agent and the reactive intermediate group are typically susceptible to hydrolysis.

8 months ago
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Protocol for conjugation of NHS–PEGn–Maleimide to Amine Particles 

An alternative method for coupling thiol-containing proteins or antibodies to amine particles is to use a heterobifunctional crosslinker containing an amine-reactive NHS ester at one end and a thiol-reactive maleimide group on the other end. NHS–PEGn–maleimide crosslinkers are available in a number of spacer lengths depending on the size of the polymer chain in the PEG component 

8 months ago
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Protocol for conjugation to Amine Particles using N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP) 

The crosslinker SPDP can be reacted with amine particles to create thiol-reactive pyridyl disulfide groups on the surface. Thiol-containing proteins or other thiol molecules can be reacted with these activated particles to result in disulfide linkages, which are reversible by reduction

8 months ago
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Protocol for conjugation of biomolecule to amine particles using two-step coupling method via glutaraldehyde

Glutaraldehyde-mediated immobilization onto amine particles can be performed with some control using two main reaction routes.

8 months ago
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General Protocol for Conjugating Biomolecules to Amino-Modified Beads

This protocol outlines a general method for covalently conjugating biomolecules to amine-modified beads. It uses a homobifunctional crosslinker, such as glutaraldehyde, imidoesters, or NHS esters, to link available amine groups on a ligand to the amine-functionalized beads.

8 months ago
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General Protocol for Coupling Biomolecules to Carboxylate Particles using EDC/Sulfo-NHS

Carboxylate particles can be coupled to amine-containing molecules using a number of reaction strategies. The most frequently used method involves an aqueous two-step coupling process using EDC and NHS or sulfo-NHS to form an amide bond with a protein or other molecules.

This protocol describes a general method for covalently conjugating amine-containing biomolecules (e.g., proteins, antibodies, oligonucleotides) to carboxylated particles, utilizing EDC (1-Ethyl-3-[3-dimethylaminopropyl]carbodiimide) and Sulfo-NHS (N-hydroxysulfosuccinimide) chemistry.General Protocol for Coupling Biomolecules to Carboxylate Particles using EDC/Sulfo-NHS

8 months ago
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