Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1995-01-30
1998-11-17
Scheiner, Toni R.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 2, 435975, 436 63, 436 64, 436514, 436515, 436813, 436826, G01N 33574, G01N 3353, G01N 3348, A01N 102
Patent
active
058374744
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
Field of the Invention
The present invention pertains to a method of detecting malignant tumors and of determining their tumor mass through the characteristics of the serum-immune globuline G (IgG) of the patient.
It has been known for several years that in patients with malignant tumors one can detect a significant change in the amounts of the sulfhydryl or disulfide groups (measured by means of the dithionitrobenzoate reaction) within the serum-IgG. The experimental measurement value for the ratio of the sulhydryl to the disulfide groups is denoted the .SIGMA.S-value; whereby the reason for the change in the .SIGMA.S-value was unknown until recently.
In the course of further research in this area it has been shown that the change of the .SIGMA.S-value can be traced to a shift in the ratio of the subclass IgG1 relative to the other subclasses (e.g. IgG2).
This interrelation is new and, besides fundamental scientific knowledge, it offers the person of skill in the art a possibility of utilizing the phenomenon in diagnostics.
Independently thereof, research in this field has repeatedly shown that the measurement values in fresh blood or serum samples change relatively strongly within a short period of time. Performing the various analysis procedures for IgG on untreated blood or serum samples, therefore, does not render reproducible and plottable results.
SUMMARY OF THE INVENTION
It is therefore an object of the invention to provide results, reproducible during a sensible period of time of at least one day, with regard to the possible presence and the size and the development of a malignant tumor in a patient.
This is attained, in accordance with the invention, in that the fresh blood or serum samples of the patient are stabilized with radical inhibitors, preferably peroxide dimutase (SOD), and in that the shift is determined in the serum of the samples of the ratio of at least one of the IgG subclasses relative to the sum of the IgG subclasses as compared to the normal ratio.
Preferably, the shift of the ratio of the subclass IgG1 and/or the subcass IgG2 is determined in the serum of the stabilized samples relative to the sum of the IgG subclasses as compared to the normal ratio.
Examination performed on untreated samples with regard to the stability of the IgG1 values with the use of the radial immuno diffusion process (RID) according to Mancini exhibited a strong loss of the serum IgG1 level when the serum samples stand exposed to air. This effect ensues virtually immediately after blood withdrawal and it averages 20% after seven hours, at least 25% after 36 hours, whereby determinations are made with blood samples of healthy donors of both sexes.
The extent of the effect depends, among other things, on the level of the initial value: Samples with a high IgG1 value usually show a stronger reduction of the value, but it appears as though, according to preliminary results, other factors may be involved as well.
While the foregoing makes it already clear that an exact determination of the IgG1 level in the blood serum is not possible in the clinical environment without an effective stabilization, such stabilization becomes an absolute necessity when the shift in the ratio among the IgG subclasses is to be evaluated in the blood of cancer patients as a tumor marker in the spirit of the method according to the invention.
The stabilization with radical inhibitors appears to be very promising, but clearly the best results have so far been achieved with the addition of SOD. The SOD may be added to the fresh complete blood samples or to the serum samples after the blood cell material has been separated out.
Very advantageous stabilization results are attained when the stabilization of the serum with SOD is performed at a mass ratio of 100-1,000, preferably 300-400 iU SOD per ml serum.
It is thereby advantageous to add the SOD to the serum samples in solution with a concentration of 5,000-10,000 iU SOD/ml. The samples which are stabilized in this manner are subsequently tested wi
REFERENCES:
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patent: 4757002 (1988-07-01), Joo
patent: 5227405 (1993-07-01), Fridovich et al.
Fluorescence Changes in Human Gamma-Globulin Induced . . . , D.G. Wickens et al., 1983, Elsevier Biomedical Press, pp. 607-616.
Schauenstein et al. "Labile disulfide bonds and free thiol groups in human IgG II. Characteristic changes in malignant diseases corresponding to shifts of IgG1 and IgG2 subclasses" Int. Archs. Allergy appl. Immuno. 80, pp. 180-184, 1986.
Smola et al "SS, a measure of reactive sulfur groups of immunoglobulin G, is a sensitive tumor marker discriminating different stages of breast cancer" Cancer vol. 68, pp. 1026-1030, 1991.
Khanna et al "Serum immunoglobulins in squamous cell carcinoma of the oral cavity" J. of Surg. Oncology, vol. 20, pp. 46-48, 1982.
European Patent Office, Database WPI, Section Ch, Week 7239, Derwent Publications Ltd, London, GB; Class A03, AN 72-62480T & JP-47 037530 (Yamada).
Dachs Franz
Schauenstein Erwin
Schauenstein Konrad
Eyler Yvonne
Greenberg Laurence A.
Lerner Herbert L.
Scheiner Toni R.
Stemer Werner H.
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