Multiplex IHC – Tech Talk

Multiplex IHC

Biocare™ Medical is the proven leader in providing Multiplex antibody cocktails and detection platforms that enable simultaneous IHC staining with multiple antibodies on a single slide. Our Multiplex detection system simplifies procedures by conserving precious tissue, saving valuable technician time, reducing the number of reagents and steps in the protocol, and allowing for simultaneous evaluation of multiple targets on one slide.

Prostate cancer stained with CK HMW + p63 + AMACR (RM)

Antibody Cocktails

Early on, Biocare recognized the market need for evaluating morphologically distinct markers to aid in solving complex clinical problems and simplifying interpretation, all while conserving precious patient tissue.

The most glaring clinical need was determined to be the ability to differentiate between prostatic intraepithelial neoplasia (PIN) and carcinoma of the prostate. There also needed to be a way to clearly identify any microinvasion or micrometastasis into adjacent prostate tissue. Additionally, this would need to be performed in a clinical laboratory, with superior accuracy and specificity. All of this whilst conserving the limited tissue (prostate needle biopsies are thin filaments of tissue) and reducing the time to result.

With these constraints and needs in mind, Biocare developed our flagship Multiplex IHC cocktail: CK HMW + p63 + AMACR (RM) [formerly known as PIN-4]. Studies have shown that combinations of CK HMW [34βE12], p63, and/or AMACR may be useful in the evaluation of normal prostate glands, prostatic intraepithelial neoplasia (PIN) and prostatic adenocarcinoma1,2,3. In prostate, CK HMW [34βE12] has been shown to be a useful marker of basal cells of normal glands and PIN, a precursor lesion to prostatic adenocarcinoma; whereas invasive prostatic adenocarcinoma typically lacks a basal cell layer3-5. p63, a homolog of the tumor suppressor p53, has been detected in nuclei of the basal epithelium in normal prostate glands; however, it was not expressed in malignant tumors of the prostate6. α-Methylacyl coenzyme A racemase (AMACR), also known as P504S, has been shown to be a specific marker of prostatic adenocarcinoma7-10. Additionally, prostate glands involved in PIN have been found to express AMACR, whereas AMACR was nearly undetectable in benign glands10-11.

  • Conserve tissues, time and money = Cost-effective
    • A single Multiplex IHC can replace up to 5 single Ab stains; reducing labor and reagent costs by at least 50%
  • Accurate analysis and easy interpretation of staining results
  • Increase predictive value by combining highly sensitive & highly specific antibodies on one slide
  • More rapidly exclude or diagnose disease states
  • Eliminate multiple slides to evaluate antigen ratios

Multiplex IHC Products

We have also developed Multiplex IHC antibody cocktails for additional tissue applications: Breast, Bladder, Lung, Skin, Colon, Lymphoma, Tumors of Unknown Origin, and Cell Death vs. Proliferation.

Tissue Multiplex Product
Prostate CK HMW + p63 + AMACR (RM)
Breast CK5/14 + p63 + CK7/18
Bladder Uro-2
Lung Desmoglein 3 + p40 (M) + Napsin A (RM)
Skin Pan Melanoma + Ki-67
Colon CDX2 (M) + CDH17 (RM)
Lymphoma Kappa (M) + Lambda (P)
Tumor of Unknown Origin CDX2 + CK7
Death vs. Proliferation Ki-67 + Caspase 3

Clinical References:
1. Shah RB, et al. Am J Clin Pathol. 2004 Oct; 122(4):517-23. 2. Sung MT, et al. Hum Pathol. 2007 Feb; 38(2):332-41. 3. Bostwick DG, Qian J. Mod Pathol. 2004 Mar; 17(3):360-79. 4. Humphrey PA. J Clin Pathol. 2007 Jan; 60(1):35-42. 5. Shah RB, et al. Am J Surg Pathol. 2002 Sep; 26(9):1161-8. 6. Signoretti S, et al. 2000 Dec; 157(6):1769-75. 7. Xu J, et al. Cancer Res. 2000 Mar 15; 60(6):1677-82. 8. Rubin MA, et al. JAMA. 2002 Apr 3; 287(13):1662-70. 9. Luo J, et al. Cancer Res. 2002 Apr 15; 62(8):2220-6. 10. Zhou M, et al. Am J Surg Pathol. 2002 Jul; 26(7):926-31. 11. Wu CL, et al. Hum Pathol. 2004 Aug; 35(8):1008-13.