Trastuzumab quantification in serum: a new, rapid, robust ELISA assay based on a mimetic peptide that specifically recognizes trastuzumab.
Trastuzumab, a humanized monoclonal antibody directed against the epidermal growth factor receptor 2 (HER2), is a milestone in the treatment of HER2-overexpressing breast cancer patients. An enzyme-linked immunosorbent assay (ELISA) for trastuzumab has been developed for routine use in the laboratory to support clinical and pharmacokinetic studies to optimize therapy. The method relies on an antigen peptide linked to a 96-well plate via the streptavidin/biotin system. The peptide sequence mimics the extracellular portion of the HER2 receptor that is recognized by trastuzumab. The calibration range of the assay is 10 to 360 ng/mL per well, corresponding to a trastuzumab serum concentration from 5 to 180 μg/mL with a lower limit of quantification of 10 μg/mL. Validation results demonstrate that trastuzumab can be accurately and precisely quantified in human serum using this assay. The procedure was also tested in sera obtained from breast cancer patients to evaluate trastuzumab serum levels, confirming the applicability of method that could be a valid assay to use in daily laboratory practice.
Development of Innovative Therapies Unit, IRCCS AOU San Martino-IST, 16132, Genoa, Italy.
Barbara
B
Cardinali
Gianluigi
G
Lunardi
Enrico
E
Millo
Andrea
A
Armirotti
Gianluca
G
Damonte
Aldo
A
Profumo
Stefania
S
Gori
Giuseppina
G
Iacono
Alessia
A
Levaggi
Lucia
L
Del Mastro
D061067:Antibodies, Monoclonal, Humanized / Q000097:blood* / Q000276:immunology / Q000627:therapeutic use*; D000941:Antigens / Q000737:chemistry / Q000276:immunology; D001943:Breast Neoplasms / Q000097:blood* / Q000188:drug therapy / Q000473:pathology; D002138:Calibration; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D057230:Limit of Detection; D018716:Molecular Mimicry; D010455:Peptides / Q000276:immunology* / Q000378:metabolism; D011786:Quality Control; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology / Q000378:metabolism; D015203:Reproducibility of Results; D000068878:Trastuzumab
D061067:Antibodies, Monoclonal, Humanized; D000941:Antigens; D010455:Peptides; D018719:Erb-b2 Receptor Tyrosine Kinases; D000068878:Trastuzumab; C508053:ERBB2 protein, human