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HAMA ELISA Kit (Human Anti-Mouse Antibody)

INTENDED USE

HAMA ELISA kit is designed for measuring in vitro quantitative amounts of human anti-mouse antibody (HAMA) in human serum samples only. This assay has a minimum detection limit of 2.0 ng/ml.

BACKGROUND

Human anti-mouse antibody (HAMA) is present in human serum, this is because of an immune reaction that is caused by an exposure to mice or other similar agent that can induce an immune system leading to the generation of antibody which are able to bind to mouse IgG. For many decades and only until recently, mice have been used extensively in the production of monoclonal antibodies (MAbs). One main problem this can create is that patients often reacted to the mouse antibodies as if they were a foreign substance and this results in the creating a new set of antibodies directed against the mouse antibodies. This process is often more commonly known as the “HAMA-response”.

There are many examples where HAMA can even cause a decrease the effectiveness of the treatment or in some cases can create a future reaction. Anywhere from one-third to even more than half of patients that receive mouse MAbs tend to develop some form of HAMA-response and there are more than 10% of the general population which are estimated to carry some form of antibody to animal-derived antigens. Finally, it has been known to result in falsely elevated levels of a number of assays and also false negatives in others, therefore making it harder to make an accurate diagnosis. It is highly recommended that baseline levels are initially determined prior to the initiation of therapy when using murine-derived proteins.

CONTENT

All reagents supplied  need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

  • Microwell Plate: Breakable wells coated with murine IgG antigen.
  • HAMA Tracer Antibody: Consisting of HRP conjugated to murine IgG in a stabilized protein matrix.
  • Tracer Antibody Diluent.
  • Assay Buffer.
  • Wash Solution (30x Concentrated).
  • TMB HRP Substrate.
  • Stop Solution.
  • Standards (0-5): Concentration 0, 40, 135, 450, 1,500.0 ng/ml.
  • Controls 1 and Control 2.

SENSITIVITY

The minimum detection sensitivity level human anti-mouse antibody using current HAMA ELISA kit was 2.0 ng/ml. The dynamic range for this assay is 40.0 – 1,500.0 ng/ml.

ASSAY CHARACTERISTICS

– Expected Values: 25 ng/ml cut-off (99% confidence).
– Sensitivity: 2 ng/ml
– Intra Assay Precision: 5.1%
– Inter Assay Precision: 5.8%
– Linearity: 90 – 113%
– High Dose Hook Effect: None up to 1,000, 000 ng/ml

REFERENCES

  1. Value of HAMA–determination in clinical practice–an overview. Anticancer Res. (1997) 17 (4B): 3055-8. Review. Hasholzner U., et al.
  2. Aberrantly elevated TSH level due to human anti-mouse antibodies (HAMA) interference with thyrotropin assay. J S C Med Assoc. (2012) 108 (1): 12-3. Hull B.
  3. Development of ELISAs for quantification of HMFG1-specific human anti-mouse IgG and IgM antibodies. Int J Biol Markers. (2007) 22 (3): 167-171. Oei A.L., et al.
  4. Human anti-mouse antibodies: pitfalls in tumor marker measurement and strategies for enhanced assay robustness; including results with Elecsys CEA. Anticancer Res. (2000) 20 (6D): 5249-52. Nussbaum S. and Roth H.J.

ADDITIONAL INFORMATION

  • Full Name: HAMA ELISA Kit (Human Anti-Mouse Antibody)
  • Reactivity: Human
  • Sample Type: Serum
  • Sensitivity: 2.0 ng/ml

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