Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1994-04-07
1996-11-26
Ceperley, Mary E.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 795, 435 4052, 436 63, 436175, G01N 33567
Patent
active
055784526
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The invention concerns immunohistochemical staining of aldehyde-fixed and embedded tissue sections.
BACKGROUND
Tissue sections obtained from clinical specimens or animal experimentation frequently are fixed, embedded, and stored in a form suitable for later examination by light microscopy. Immunological reagents, especially monoclonal antibody reagents, currently permit examination of at least certain of these fixed tissue samples for the presence of particular antigenic compounds. Antigens of interest may be associated with a disease process or pathology, or may identify a particular cell type or tissue. In the case of recently prepared biopsy and autopsy samples, such immunohistochemical analyses are of immediate diagnostic value.
However, immunohistochemical analyses of tissue specimens have been hampered because of antigenic loss during specimen fixation. Traditional fixation methods frequently have employed aldehyde fixatives, which fix the tissue by causing cross-linking reactions within and between tissue proteins.
Two types of cross-linking reactions have been recognized. The first is a Schiff's base-type polymerization: formaldehyde condenses with the amino groups of the protein, resulting in the Schiff's base intermediate, which is capable of undergoing rapid polymerization leading to cross-linking of the proteins.
In the second type of reaction, called the Mannich reaction, the formaldehyde can react with both an amino group and an active hydrogen group, resulting in the formation of a Mannich base. Polymerization of the Mannich bases results in protein cross-linking.
Cross-links preserve tissue morphology and integrity, harden the tissue for slicing, and inhibit microbial attack. Unfortunately, the cross-linking process also causes loss of tissue antigenicity, a result which impedes the usefulness of immunological reagents on tissues fixed with aldehyde reagents such as formaldehyde. The chemistry of the cross-linking of amino acids and proteins by formaldehyde is described in Harlan and Feairheller, "Chemistry of the Cross-Linking of Collagen During Tanning," and Kelly, et al. "Cross-Linking of Amino Acids By Formaldehyde," (1976). The role of Mannich-type reactions in cross-linking of protein amino groups and aromatic amino acids with formaldehyde is discussed in Fraenkel-Conrat, et al., J. Biol. Chem. (1947) 168:99-118, and Fraenkel-Conrat and Olcott, J. Biol. Chem. (1948) 174:827-843. Further discussions of aldehyde cross-linking reactions are found in Fox, J. Histochem. Cytochem. (1985) 33:845-855; Jones, "Reactions of aldehyde with unsaturated fatty acids during histological fixation," in Fixation in Histochemistry, P. J. Stoward, ed. (1973); and Kunkel et al., Mol. Cell. Biochem. (1981) 34:3. Mannich type reactions are described in general in March, "Advanced Organic Chemistry," particularly at 333,424, 670-672 (1968).
In an attempt to circumvent the disadvantages of aldehyde fixation, alternative fixation methods have been developed, such as microwave heating (Mayers, J. Clin. Pathol. (1970) 28:273; Hopwood et al., Histochem. J. (1984) 16:1171) and alcohol immersion (Battifora and Kopinski, J. Histochem. Cytochem. (1986) 34:1095). Despite some advantages of alternative fixation methods, they have not displaced aldehyde fixation in general use. Their limited acceptance may reflect drawbacks present in these alternative methods. For example, microwave heating lyses red cells and disrupts membrane lipids. Although ethanol fixation is reported to produce improved antigenicity of tissue samples, ethanol causes increased cellular shrinkage (Battifora and Kopinski, id.) Consequently methods for restoring antigenicity to aldehyde fixed tissues continue to be useful for specimens generated by current clinical practices.
In addition, methods for restoring antigenicity are useful because of the vast number of aldehyde fixed tissue samples already in collections. These stored tissue samples provide a rich reservoir of material for retrospective immunohistochemical examination. I
REFERENCES:
patent: 5188834 (1993-02-01), Grimm et al.
P. Rumph et al., Anat. Histol. Embryol., vol. 15, No. 3, pp. 269-276 (1986).
P. Rumph et al. Anat. Histol. Embryol., vol. 17, No. 3, pp. 226-231 (1988).
A. Ross, J. Electron Microsc. Tech., vol. 5, No. 1, pp. 81-90 (1987).
E. Gendler, English Abstract of U.S.S.R. Patent No. 138,336 (1960).
K. Yasuda, HCAPLUS Abstract of Saibo, vol. 15, No. 4, pp. 545-547 (1983).
N. Yamamoto et al. HCAPLUS Abstract of Kitasato Igaku, vol. 11, No. 1, pp. 9-17 (1981).
Harlan and Feairheller, "Chemistry of the CrossLinking of Collagen During Tanning", Adv Exp Med Biol (1977) 86A:425-440.
Kelly, et al. "Cross-Linking of Amino Acids By Formaldehyde Preparation and Carbon-13 NMR Spectra of Model Compounds", Adv Exp Med Biol (1977) 86A:641-647.
Fraenkel-Conrat, et al., (1947) "The Reaction of Formaldehyde with Proteins (IV) Participation of Indole Groups. Gramicidin" J. Biol. Chem., 168:99-118.
Fox, (1985) J. Histochem. Cytochem. 33:845-855.
Jones, (1973) "Reactions of aldehyde with unsaturated fatty acids during histological fixation" Fixation in Histochemistry, P. J. Stoward, ed.
Kunkel et al., (1981) Mol. Cell. Biochem. 34:3.
March, (1968) "Advanced Organic Chemistry," particularly at 333, 424, 670-672.
Mayers, J. Clin. Pathol. (1970) 28:273.
Hopwood et al., Histochem. J. (1984) 16:1171.
Battifora and Kopinski, J. Histochem. Cytochem. (1986) 34:1095-1100.
Huang, et al., J. Lab Invest. (1976) 35:383-390.
Leong, et al., J. Pathology (1988) 156:275-282.
Shi et al., (1991) "Antigen Retrieval in Formalin-fixed, Paraffin-embedded Tissues: An Enhancement Method for Immunohistochemical Staining Based on Microwave Oven Heating of Tissue Sections" The Journal of Histochemistry and Cytochemistry, 39:741-478.
Carey et al., (1983) Advanced Organic Chemistry, 2nd ed. 58-62.
Fraenkel-Conrat et al., (1948) "Reaction of Formaldehyde with Proteins (IV) Cross-linking of Amino Groups with Phenol, Imidazole, or Indole Groups" J. Biol. Chem., 174:827-843.
Kalra Krishan L.
Malhotra Nagesh
Shi Shan-Rong
Su Sheng-Hui
Tandon Atul K.
Biogenex Laboratories
Ceperley Mary E.
LandOfFree
Enhancement of immunochemical staining in aldehyde-fixed tissues does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Enhancement of immunochemical staining in aldehyde-fixed tissues, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Enhancement of immunochemical staining in aldehyde-fixed tissues will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1972123