Exceeding Expectations in Protein and Antibody Biopharmaceutical Separations

Therapeutic proteins and monoclonal antibodies form the largest part of the growing biologics drug market and has transformed the biotechnology and biopharmaceutical industry in the last decade. Protein drugs include blood factors (Factor VIII and Factor IX), thrombolytic agents (tissue plasminogen activator), hormones (insulin, glucagon, growth hormone, gonadotrophins), hematopoietic growth factors (erythropoietin, colony stimulating factors), interferons (interferons-α, -β, -γ), interleukin-based products (interleukin-2) and additional products like tumor necrosis factor and therapeutic enzymes.

Monoclonal antibody treatment are the most effective method for diagnosis and treatment of a broad range of diseases including autoimmune, cardiovascular, infectious diseases, cancer, and inflammation with key advantages offering reduced toxicity and side effects with high targeted specificity.

Dionex provides intelligent HPLC platforms and innovative column chemistries that address every facet of protein and antibody therapeutics R&D and QA/QC challenges to ensure safe biopharmaceuticals.

Learn about our Protein and Antibody Therapeutics Applications Notebook, or browse our applications under the Related Literature tab above.

Challenges in Development for Protein and Antibody Therapeutics

Protein and antibody molecules are large with complex post-translational modifications and can only be produced via genetic engineering in living cells. As a result, the development and manufacturing of these drugs is extensive and challenging.

Complete Characterization for Safe and Efficient Biopharmaceuticals

Protein and antibody drugs may be modified after synthesis naturally (through posttranslational modifications – PTMs), through degradation processes or through human engineering. Naturally occurring modifications are inherent problems of these large molecules. Common modifications include mutations, glycosylations, oxidations, phosphorylations, amino terminal modifications (e.g. pyroglutamate), incomplete processing of the C-terminus, and asparagine (Asn) deamidations. Slight changes in cell lines and tissue culture conditions can result in changes in the protein structure and its glycosylations pattern.

During the development and production of these products, detection, characterization, and quantification of impurities, structural variants and modifications, as well as monitoring stability, are regarded as essential for demonstrating safety and efficacy of these drugs and is required by the FDA and other regulatory agencies. Common analyses tasks include:

Purity

The recombinant protein and monoclonal antibody therapeutics should be as free as possible of contaminants like non-Ig contaminants.

Specificity and Potency

Assessment of biological activity should be performed to demonstrate the efficacy of the product e.g. Binding of the MAb to the target antigen is specific.

Structural Integrity

Protein therapeutics manufacturers are required to provide complete information on primary, secondary and tertiary structure information including any higher order structure which may be acquired by various biochemical and analytical means. Typical structural information required includes:

• Fragmentation
• Aggregation
• Carbohydrate structure
• Post-translational modifications
  • Chemical degradation (deamidation, oxidation)
  • Enzymatic degradation (lysine processing, clipping)
  • Post-translational modifications (glycosylation, folding, disulfide formation)
  • Structural modifications (denaturation, aggregation)
  • Charge variants (deamidation, pyroglutamate formation, lysine processing
  • Conjugation (PEGylation, cytotoxic chemicals)