Megamolecule Synthetic Antibodies
- 技术优势
- • Modular synthesis permits the creation of molecules too large or complex to prepare using standard protein engineering/expression as such methods require polypeptides to be expressed as one or more chains in culture. Correct / folding and assembly and purification of larger molecules becomes more difficult as the complexity of the molecule is increased thereby decreasing yield and increasing cost. • The chemical linking strategy enables an efficient method to produce precisely-defined protein scaffolds of variable stoichiometry, orientation, and geometry. These attributes can be systematically altered by encoding diversity into the small molecule linkers and by the order of attachment. Current protein engineering methods are limited to the intrinsic repertoire of natural polypeptide folds / peptide linkers to achieve a desired construct. Therefore, non-natural formats that may exhibit greater efficacy cannot be prepared using conventional engineering methods. • The mild, rapid, and site-specific nature of the linking reactions allows for diverse effector molecules to be attached to an antibody fragment in precise stoichiometry in high yield and away from the antibody-like fragment. Current chemical methods require the engineering of a reactive amino acid or the use of a preexisting reactive amino acid side chain to attach payloads to the antibody scaffold. In many cases, products of these reactions yield heterogeneous populations of products. Therefore, such reactions must be arduously optimized prior to scale-up. Such direct amino-acid modifications may also not be stable or cause deleterious effects to the pharmacological properties of the parent molecule (e.g. increased hydrophobicity, aggregation, immunogenicity, etc.) therefore a great deal of effort must be directed toward choosing a suitable location for attaching the payload. • The modularity of the linking reactions also enables the joining of effector molecules (e.g. toxins, glycosylated proteins) that cannot be produced in one culture system efficiently due to toxicity constraints or metabolic profiles of the expression hosts.
- 详细技术说明
- A novel method to join antibody fragments to produce megamolecule antibody like drugs #therapeutics #antibody
- *Abstract
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BACKGROUND
Biologic drugs, specifically monocolonal antibodies (mAbs) and those that incorporate antibody fragments were responsible for ~$75B of total pharmaceutical sales in 2013. Due to their ability to specifically target disease-related proteins in circulation or on cell surfaces, industry has put a great deal of effort in creating 'next-generation antibody therapeutics' – those that show enhanced therapeutic potential – by increasing their valency, attaching effector molecules that increase cytotoxicity and half-life, and by using formats that direct the immune system toward clearing targets. These molecules, however, are often challenging to produce in high-yield and with synthetic flexibility that permits a desired function to be systematically optimized.
ABSTRACT
Northwestern researchers developed a novel method of joining antibody fragment and antibody-like fragment-fusion proteins in a modular fashion using small molecule linkers that react site specifically in high yield with rapid kinetics under mild reaction conditions. This method permits not only the scalable preparation of molecules with molecular weights >250 kDa, but also allows for the creation of an incredibly diverse library of antibody-like drugs with altered valency, geometry, effector function, and stoichiometry simply by the combination of a comparatively smaller number of building blocks.
- *Inventors
- Milan Mrksich* Justin Modica
- *Publications
- Modica, J. A., Skarpathiotis, S. and Mrksich, M. (2012), Modular Assembly of Protein Building Blocks To Create Precisely Defined Megamolecules. ChemBioChem, 13: 2331–2334. doi:10.1002/cbic.201200501
- 国家/地区
- 美国
