Advanced Techniques in Analytical Chemistry

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Questions and Answers

Which technique is NOT mentioned as a method used in the quality control of biotechnologically produced drugs?

Size Exclusion Chromatography

Binding assays

Protein sequencing using the Edman reaction

High-performance liquid chromatography (correct)

What is the physical source of human growth hormone?

Porcine thyroid

Salmon

Human pituitary gland (correct)

Synthetic production

Which drug is used for the reversal of insulin-induced hypoglycemia?

Immunoglobulins

Human growth hormone

Glucagon (correct)

Insulin

What is the therapeutic action of vaccines mentioned in the text?

Stimulation of immune system to develop B-lymphocyte memory cells Signup and view all the answers

Which technique is potentially many times more efficient than HPLC in separating power?

Capillary electrophoresis Signup and view all the answers

In which technique do all proteins pick up the same proportion of SDS on a w/w basis, resulting in identical negative charge regardless of their size?

SDSPAGE Signup and view all the answers

Which technique uses a gel prepared with a pH gradient across it for separation based on the pI values of the proteins?

Isoelectric focusing Signup and view all the answers

Which technique separates molecules based on molecular size, with larger molecules eluting first?

Size Exclusion Chromatography (SEC) Signup and view all the answers

Which type of binding assay uses an antibody to the protein drug and quantifies the amount of antibody bound by measuring enzyme activity or radioactivity?

Enzyme-linked immunosorbent assay (ELISA) Signup and view all the answers

What is a downside of binding assays such as ELISA and RIA?

Increased cost Signup and view all the answers

Which method is used for the production of Humira (adalimumab) by AbbVie?

CHO production method Signup and view all the answers

What is the principle of Capillary Electrophoresis (CE)?

Applies a high potential to a fused-silica capillary tube filled with mobile phase Signup and view all the answers

What influences ion migration in Capillary Electrophoresis (CE)?

Electric field and buffer pH Signup and view all the answers

What is the equation for analyte's ion velocities (me) in an electric field in Capillary Electrophoresis (CE)?

$me = q/6\text{phr}$ Signup and view all the answers

What are some popular indications for Keytruda (pembrolizumab) by Merck?

Melanoma and non-small-cell lung cancer Signup and view all the answers

What does Capillary Electrophoresis (CE) separate based on?

Analyte's ion velocities in an electric field Signup and view all the answers

Which drug is produced using the mouse myeloma cells production method?

Remicade (infliximab) by Johnson & Johnson Signup and view all the answers

What does Capillary Electrophoresis (CE) control through instrumentation?

Migration time and dispersion Signup and view all the answers

Which drug has indications for Crohn's disease, ulcerative colitis, and rheumatoid arthritis?

Remicade (infliximab) by Johnson & Johnson Signup and view all the answers

Which technique does Capillary Electrophoresis (CE) use to separate enantiomers, impurities, and drugs?

Electric field to separate enantiomers, impurities, and drugs. Signup and view all the answers

Study Notes

Hepatitis A: A contagious disease caused by the Hepatitis A virus, leading to inflammation and damage to the liver.
Proteinaceous drugs: Drugs that consist of proteins, some of which are among the best-selling in the market.
Some popular protein drugs and their features:
- Humira (adalimumab) by AbbVie: CHO production method, $20.2 billion sales, indications for rheumatoid and psoriatic arthritis, Crohn‘s disease, and ulcerative colitis.
- Avastin (bevacizumab) by Roche: CHO production method, $6.4 billion sales, indications for metastatic colorectal, non-small-cell lung, glioblastoma, and metastatic kidney cancers.
- Herceptin (trastuzumab) by Roche: CHO production method, $6.4 billion sales, indications for HER2-positive breast and metastatic gastric cancers.
- Remicade (infliximab) by Johnson & Johnson: Mouse myeloma cells production method, $6.3 billion sales, indications for Crohn's disease, ulcerative colitis, and rheumatoid arthritis.
- Keytruda (pembrolizumab) by Merck: CHO production method, $6.1 billion sales, indications for melanoma and non-small-cell lung cancer.
Capillary Electrophoresis (CE):
- Analyte separation technique using electric field to separate enantiomers, impurities, and drugs.
- Principle: applies a high potential to a fused-silica capillary tube filled with mobile phase, containing an aqueous component and an electrolyte.
- Analytes migrate at rates determined by charge and ionic radius, and neutral analytes also migrate due to electro-osmotic flow (EOF).
- High Precision and Selective: in drug quantification, quality control, separating enantiomers, impurity profiling, and analyzing drugs/metabolites in biological fluids.
Capillary Electrophoresis (CE) Background:
- Separation based on analyte's ion velocities (me) in an electric field, which equals the charge on the molecule (q) divided by the frictional drag (FF) of the medium (h, r, n).
- At the point of equal FE and FF (qE = -6phrn), me = q/6phr, and ions will start to migrate when FE > FF.
Capillary Electrophoresis (CE) Ion migration:
- Ion migration in CE is influenced by the EOF, causing everything to move towards the cathode, and rates vary depending on charge.
- Cations: rate = ion mobility + EOF; Neutrals: rate = EOF; Anions: rate = EOF - ion mobility.
Capillary Electrophoresis (CE) Instrumentation:
- Controls separation through migration time, dispersion, injection plug length, Joule heating, solute/wall interactions, and electro-dispersion.
- Influenced factors: buffer pH, buffer strength, temperature, electric field, and surfactant concentration.
- Surfactants: cationic can reduce EOF or reverse flow towards the anode, and anionic surfactants increase EOF.
- Coating: neutral coatings reduce EOF, while ionic coatings have marked effects on EOF.
Examples of CE applications:
- Separation of NSAIDs based on ionic radius (sulindac, indomethacin, piroxicam, tiaprofenic acid, and aceclofenac).
- Separation of human cerebrospinal fluid from patients with multiple sclerosis, cerebral infarction, and neurosis.
- Comparison of HPLC-MS and HPCE-MS for the analysis of aviptadil, a synthetic injectable formulation of human vasoactive intestinal peptide, in the treatment of acute respiratory distress syndrome, asthma, and COPD.

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Description

Learn about high-performance capillary electrophoresis and quality control methods for biotechnologically produced drugs in this advanced lecture. Understand the principles behind these techniques and their applications in biotechnological analysis.