Abstract
In diagnostic pathology and immunocytochemical research, immunohistochemical techniques using the streptavidin–biotin–peroxidase system have played an extremely valuable role. This system, based on the high affinity of streptavidin for biotin, may, however, provoke false positive results because of endogenous streptavidin-binding sites in human tissues. With the advent of the antigen retrieval procedure and signal amplification method, this problem can be serious enough to cause mistakes in interpreting immunohistochemical staining results. Therefore, we examined the distribution of endogenous biotin-like molecules in various human tissues and the influence of various antigen retrieval procedures with or without signal amplification using biotinylated tyramine to reveal these biotin-like activities. We observed that endogenous biotin-like molecules were present in a wide range of tissues, and their activity was markedly enhanced by employing antigen retrieval procedures or signal amplification. Furthermore, the extent to which the activity of endogenous biotin-like activities was enhanced depended on the kinds of antigen retrieval procedures and signal amplification employed. Pressure cooking and tyramine amplification with microwave heating showed the highest activities. These results show that the antigen retrieval procedures and signal amplification with tyramine can enhance the activity of endogenous biotin or biotin-like molecules as well as antigenicity, which can be a pitfall in the interpretation of immunohistochemical data.
Similar content being viewed by others
References
Bhattacharjee J, Cardozo BN, Kamphuis W, Kamermans M, Vrensen GF (1997) Pseudoimmunolabelling with the avidin–biotin–peroxidase complex due to the presence of endogenous biotin in retinal muller cells of goldfish and salamander. J Neurosci Meth 77: 75–82.
Bobrow MN, Harris TD, Shaughnessy KJ, Litt GJ (1989) Catalyzed reporter deposition, a novel method of signal amplification. J Immunol Meth 125: 279–285.
Bussolati G, Gugliotta P, Volante M, Pace M, Papotti M(1997) Retrieved endogenous biotin: a novel marker and a potential pitfall in diagnostic immunohistochemistry. Histopathology 31: 400–407.
Cooper KM, Kennedy S, McConnel S, Kennedy DG, Frigg M(1997) An immunohistochemical study of the distribution of biotin in tissues of pigs and chickens. Res Vet Sci 63: 219–215.
Dakshinamurti K, Mistry SP (1963) Tissue and intracellular distribution of biotin-14COOH in rats and chicks. J Biol Chem 238: 294–296.
Duhamel RC, Whitehead JS (1990) Prevention of nonspecific binding of avidin. Meth Enzymol 184: 201–207.
Fleming H, Condon R, Peterson G, Guck I, Prescott E, Chatfield K, Duff M (1998) Role of biotin-containing membranes and nuclear distribution in differentiating human endometrial cells. J Cell Biochem 71: 400–415.
Kashima K, Yokoyama S, Daa T, Nakayama I, Nickerson PA, Noguchi S (1997) Cytoplasmic biotin-like activity interferes with immunohistochemical analysis of thyroid lesions: a comparison of antigen retrieval methods. Mod Pathol 10: 515–519.
Kerstens MJ, Poddighe PJ, Hanselaar AGJM (1995) A novel in situ hybridization amplification method based on the deposition of biotinylated tyramine. J Histochem Cytochem 43: 347–352.
Levin SM, Macklin WB (1988) Biotin enrichment in oligodendrocytes in the rat brain. Brain Res 444: 199–203.
Lu CS, Kashima K, Daa T, Yokoyama S, Yanagisawa S, Nakayama I (2000) Immunohistochemical study of the distribution of endogenous biotin and biotin-binding enzymes in ductal structures of salivary gland tumours. J Oral Pathol Med 29: 445–451.
Merz H, Malisius R, Mannweiler S, Zhou R, Hartmann W, Orscheschek K, Moubayed P, Feller AC (1995) Immunomax, a maximized immunohistochemical method for the retrieval and enhancement of hidden antigens. Lab Invest 73: 149–156.
Rowly RF, Eisenberth GS (1982) Selective binding of fluoresceinatedavidin to A-cells of sectioned pancreas. Diabetes 31: 107–109.
Sickel JZ, Di Santágnese PA (1994) Anomalous immunostaining of ‘optical clear’ nuclei in gestational endometrium. A potential pitfall in the diagnosis of pregnancy-related herpesvirus infection. Arch Pathol Lab Med 118: 831–833.
Wood GS, Warnke R (1981) Suppression of endogenous avidin-binding activity in tissues and its relevance to biotin-avidin detection systems. J Histochem Cytochem 29: 1196–1204.
Wood HG, Borden R (1977) Biotin enzymes. Ann Rev Biochem 46: 386–411.
Yokoyama S, Kashima K, Inoue S, Daa T, Nakayama I, Moriuchi A (1993) Biotin-containing intranuclear inclusions in endometrial glands during gestation and puerperium. Am J Clin Pathol 99: 13–17.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hyung Kim, S., Cheon Jung, K., Kee Shin, Y. et al. The Enhanced Reactivity of Endogenous Biotin-like Molecules by Antigen Retrieval Procedures and Signal Amplification with Tyramine. Histochem J 34, 97–103 (2002). https://doi.org/10.1023/A:1020954611464
Issue Date:
DOI: https://doi.org/10.1023/A:1020954611464