Abstract
Starting from the phenomenon that the amount of circulating CEA in patients' sera did not significantly influence immunoscintigraphic visualization of CEA expressing tumors, we built up an in vitro model to explain this phenomenon. Blocking experiments in this model system showed that the CEA specific MAbs BW 431/26 and BW 431/31 could not be inhibited in their binding to cell associated CEA, if they were preincubated with a 20 molar excess of serum CEA. In contrast, the CEA-NCA cross reactive MAbs could be inhibited in their binding to tumor associated CEA under identical conditions. These data combined with western blotting analysis of patients' sera and affinity constant determinations argue that conformational changes in serum CEA cause a decreased affinity of the CEA specific MAbs to serum CEA allowing a preferential binding to tumor associated CEA.
Similar content being viewed by others
Abbreviations
- CEA:
-
carcinoembryonic antigen
- NCA:
-
nonspecific cross-reacting antigen
- DTPA:
-
diethylene triamine pentaacetic acid
- FBS:
-
fetal bovine serum
- EDTA:
-
ethylene diamine tetraacetic acid
- PCA:
-
perchloric acid
- R.T.:
-
room temperature
- PBS:
-
phosphate buffered saline
- BSA:
-
bovine serum albumin
- SDS-PAGE:
-
sodium dodecyl-sulfate polyacrylamide gel electrophoresis
References
Berson SA, Yalow RS (1959) Quantitative aspects of the reaction between insulin-binding antibody. J Clin Invest 38:1996–2016
Bosslet K (1987) Monoclonal antibodies suited for the immunoscintigraphic differential diagnosis of human lung cancer. Meeting Abstract: 3rd International Congress on Hormones and Cancer, September 1987, Hamburg, FRG
Bosslet K, Lüben G, Schwarz A, Hundt E, Harthus HP, Seiler FR, Muhrer C, Klöppel G, Kayser K, Sedlacek HH (1985) Immunohistochemical localization and molecular characteristics of three monoclonal antibody-defined epitopes on carcino-embryonic antigen (CEA). Int J Cancer 36:75–84
Bosslet K, Kern HF, Kanzy EJ, Steinstraesser A, Schwarz A, Lüben G, Schorlemmer HU, Sedlacek HH (1986) A monoclonal antibody with binding and inhibiting activity towards human pancreatic carcinoma cells. Cancer Immunol Immunother 23:185–191
Bosslet K, Schwarz A, Steinstraesser A, Kuhlmann L, Seidel L, Schulz G (1988) Immunszintigraphie kolorektaler Karzinome und spezifische Immuntherapie von Pankreaskarzinomen mittels monoklonaler Antikörper. Wiener Med Wschr 11:255–257
Boyum A (1976) Isolation of lymphocytes, granulocytes and macrophages. Scand J Immunol 5:9–15
Buchegger F, Schreyer M, Carrel S, Mach JP (1984) Monoclonal antibodies identify a CEA cross-reacting antigen of 95 kD (NCA-95) distinct in antigenicity and tissue distribution from the previously described NCA of 55 kD. Int J Cancer 33:643–649
Chandrasekaran EV, Davila M, Nixon DW, Goldfarb M, Mendicino J (1983) Isolation and structures of the oligosaccharide units of carcinoembryonic antigen. J Biol Chem 258:7213–7222
Dillman RO, Beauregard JC, Sobol RE, Royston I, Bartholomew RM, Hagan PS, Halpern SE (1984) Lack of radioimmunodetection and complications associated with monoclonal anticarcino-embryonic antigen antibody. Cancer Res 44:2213–2218
Egan ML, Engvall E, Ruoslahi EI, Todd CW (1977) Detection of circulating tumor antigens. Cancer Res 40:458–466
Engvall E, Shively JE, Wrann M (1978) Isolation and characterization of the normal cross-reacting antigen: Homology of its NH2-terminal amino acid sequence with that of carcinoembryonic antigen. Proc Nat Acad Sci USA 75:1670–1674
Ey PL, Prowse SJ, Jenkin CR (1978) Isolation of pure IgG1, IgG2a, IgG2b immunoglobulins from mouse serum using protein Asepharose. Immunochemistry 15:429–436
Fraker PJ, Speck JC (1978) Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetra chloro-3,6-diphenyl glycoluril. Biochem Biophys Res Comm 80:849–857
Fuks A, Shuster J, Gold P (1980) Theoretical and practical considerations of the utility of the radioimmunoassay for carcinoembryonic antigen in clinical medicine. In: Sell S (ed) Cancer markers, Human, Clifton NJ, pp 315–327
Gold P, Freedman SO (1965a) Demonstration of tumor-specific antigens in human colonic carcinomata by immunological tolerance and absorption techniques. J Exp Med 121:439–462
Gold P, Freedman SO (1965b) Specific carcinoembryonic antigens of the human digestive system. J Exp Med 122:467–480
Kleist S von, Chavanel G, Burtin P (1972) Identification of an antigen from normal human tissue that cross-reacts with the carcino-embryonic antigen. Proc Nat Acad Sci (Wash) 69:2492–2494
Krupey J, Gold P, Freedman SO (1968) Physicochemical studies of the carcinoembryonic antigen of the human digestive system. J Exp Med 128:387–398
Kuroki M, Arakawa F, Higachi H, Matsunaga A, Okamoto N, Takakura K, Matsuoka Y (1987) Epitope mapping of the carcinoembryonic antigen by monoclonal antibodies and establishment of a new improved radioimmunoassay system. Jpn J Cancer Res 78:386–396
Mach JP, Buchegger F, Forni M, Ritschard J, Berche C, Lumbroso J-D, Schreyer M, Girardet C, Accolla RC, Carrel S (1981) Use of radiolabelled monoclonal anti-CEA antibodies for the detection of human carcinomas by external photoscanning and tomoscintigraphy. Immunology today 2:239–249
Markwell MAK, Fox CF (1978) Surface-specific iodination of membrane proteins of viruses and eucaryotic cells using 1,3,4,6-tetrachloro-3,6-diphenylglycoluril. Biochemistry 17:4807–4817
Nichols EJ, Kannagi R, Hakomori SI, Krantz MJ, Fuks A (1985) Carbohydrate determinants associated with carcinoembryonic antigen (CEA). J Immunol 135:1911–1913
Oikawa S, Nakazato H, Kosaki G (1987a) Primary structure of human carcinoembryonic antigen (CEA) deduced from cDNA sequence. Biochem biophys Res Commun 142:511–518
Oikawa S, Imajo S, Noguchi T, Kosaki G, Nakazato H (1987b) The carcinoembryonic antigen (CEA) contains multiple immunoglobulin-like domains. Biochem biophys Res Commun 144:634–642
Rogers GI (1983) Carcinoembryonic antigens and related glycoproteins. Molecular aspects and specificity. Biochim Biophys Acta 695:227–249
Scatchard G (1949) The attractions of proteins for small molecules and ions. Ann NY Acad Sci 51:660–672
Steinsträsser A, Schwarz A (1986) Quality control of iodinated monoclonal antibodies. Nucl Med 25:A75–76
Steinsträsser A, Schwarz A, Kuhlmann L (1987) Monoclonal antibodies: Optimising of labelling procedures by quality control methods. Third European Symposium on Radiopharmacy and Radiopharmaceuticals; Kristensen K, Dige-Peterson H, Marqversen J (eds) Elsinore, Abstract, pp 68–69, Elsinore, Denmark, Isotope pharmacy bronshoj, May 1–4, 1987
Towbin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Nat Acad Sci USA 76:4350–4354
Walker WHC (1977) The scatchard plot in immunometric assay. Clin Chem 23:588–590
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bosslet, K., Steinsträsser, A., Schwarz, A. et al. Quantitative considerations supporting the irrelevance of circulating serum CEA for the immunoscintigraphic visualization of CEA expressing carcinomas. Eur J Nucl Med 14, 523–528 (1988). https://doi.org/10.1007/BF00286769
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00286769