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Quality Control And Validation

Updated June 4, 2015

Transcript

ICH Guidelines

Monoclonal antibodies are derived by recombinant DNA
Cell line should follow ICH Q5B & ICH Q5D
CHO cell line is derived from animals so production should also be subject to ICH Q5A
Q5A: guidelines concerning “viral safety evaluation of biotechnology products derived from animal cells of human or animal origin.”
These ICH guidelines were important for establishing the production quality of Herceptin and establishing what the acceptance criteria for each batch would be.

Identity – is it the right drug as indicated on the label?
Quantity– how much drug is there and is it consistent with the labeled amount?
Impurities – are there process impurities or degradation impurities?
Dissolution – does the active ingredient dissolve out of the dosage unit so that the drug is available for the body to absorb?

Active pharmaceutical ingredient in Herceptin is a monoclonal antibody
Several sections of ICH guidelines are for its development and production
Monoclonal antibodies should be fully characterized and their binding properties and specifications known
ICH Q6B lays out what needs to be known about a monoclonal antibody in order to be approved
Gives “guidance on justifying and setting specifications for proteins and polypeptides”-ICH Q6B
Guideline states characterization of monoclonal antibody should include: biological activity, purity, impurity, quantity, and immunochemical and physicochemical properties.

150 mg trastuzumab, 3.36mg L - Histidine HCl, 2.16 mg L-Histidine, 136.2mg Trehalose dihydrate, and .6 mg Polysorbate 20

Estimated 231,840 new cases invasive breast cancer to be diagnosed in women in the U.S. in 2015,
60,290 new cases of non-invasive breast cancer.
2,350 new cases invasive breast cancer expected in men in 2015
Man’s lifetime risk of breast cancer is about 1 in 1,000
Breast cancer is the 2nd most commonly diagnosed cancer among American women
Estimated that about 30% newly diagnosed cancers in women will be breast cancers

•Lumps or masses in the breast

•Swelling part(s) of a breast

•Skin irritation or dimpling

•Breast or nipple pain

•Nipple retraction (inverted nipple)

•Nipple discharge other than when pregnant

-scores 0-3

-HER2+ scores a 3

Appearance
Color, opalescence,& clarity must match reference sample
Herceptin powder comes as white to pale yellow
Opalescence and clarity can be assayed using a spectrophotometer
Identity
sequence and binding structure
Peptide Mapping by RP-HPLC
reverse phase - high performance liquid chromatography
cleave peptide bonds
chromatographic separation of the peptides
analysis done in parallel with the reference
confirms that no changes to the primary structure have taken place
MALDI (matrix assisted laser desorption/ionization) Mass Spectrometry
analyte is put into a crystal matrix which causes the molecules to ionize
laser then hits the matrix and the energy transfer causes the molecules to enter gas phase
high electric field, causes it to propel into a flight tube where a detector sequences them

Purity
free of unwanted substances
PAGE (Polyacrylamide Gel Electrophoresis)
uses polyacrylamide as the matrix for electrophoresis
treated with sodium dodecyl in order to make it negative
stained, purity would be indicated by bands matching those or the reference material
IEC (ion exchange chromatography)
rate of passage through the column is determined by particle attraction
results should match the reference sample
Safety
endotoxin, pyrogen, microbe free
LAL (limulus amebocyte lysate) method
amebocytes from horseshoe crab blood used to detect pyrogens and endotoxins
three types of LAL: chromogenic, turbidimetric, gel clot