Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Utility Patent
1998-11-23
2001-01-02
Swartz, Rodney P. (Department: 1641)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C422S051000, C422S051000, C422S051000, C422S082110, C435S007200, C435S007300, C435S007320, C436S172000, C436S518000, C436S524000, C356S340000, C356S937000, C359S580000, C359S586000, C359S589000
Utility Patent
active
06168921
ABSTRACT:
BACKGROUND OF THE INVENTION
The invention relates to a method and system for measuring the binding reactions of atoms or molecules to other atoms or molecules which are locked to a surface.
A quantitative and qualitative acquisition of atoms and molecules is becoming nowadays more and more important. Particular in medical technology, biotechnology and pharmaceutical technology such methods are for example employed for the acquisition of antigen/antibody complexes on solid or fluid phases. For in vitro diagnosis of various diseases, one uses the specific bonding characteristics of antibodies to their antigens. The analysis, in particular ascertaining certain molecules and the quantitative determination of these molecules, gives information for example on the immune status, the course of a disease, the deficiency or excess of body-specific substances, defects in genetic material and much more.
Known methods of this type at the present time function such that with the help of physical and/or chemical methods, antibodies or antigens for example of complex material mixes are selectively bound and then are directly or indirectly detected or quantified. Examples for this are the determining of immune globulins in body fluids, determining antibodies to infectious diseases (IGG, IGA, IGM), determining antibodies involved in allergies (IGE, IGG, IGM), determining auto-immune antibodies involved in immune diseases, for example, rheumatism, determining proteins and peptides, determining polyclonal and monoclonal antibodies, hormones, determining breakdown products or secretions for example as disease indicators, determining substances foreign to the body, e.g medication, drugs, environmental chemicals.
The methods currently employed in medical technology are apparatus-extensive, time consuming, partly highly complicated, and with regard to their discrepancy, have certain risks. These known methods work essentially according to the principle that firstly molecules which can selectively bind other molecules or atoms, for example antigens, may be bound onto a solid phase as a substrate. After an incubation with which all molecules concerned are bound onto the solid phase, the excess or non-bound material is removed by way of one or more washing steps. Then this solid phase, adhered with molecules for selective bonding (for example, antigens) is exposed to the sample to be examined.
According to the lock-and-key principle, the molecules or atoms located in the sample (for example, antibodies) are bound to the molecules/antigens of the solid phase, in as far as they are present (primary bonding). After an incubation, after which all molecules concerned are bound, then again the removal of the excess non-bound material is effected in one or ore washing steps. Where appropriate, then for strengthening, a further antibody of animal origin is added, which is directed against the substance in question. After the incubation has been effected then, again, the removal of excess non-bound material is effected in one or more washing steps.
Finally, by way of the addition of previously marked molecules (e.g., by way of enzymes, radioactive substances, light emitting substances or likewise), a qualitative or quantitative detection of primary bondings is effected by activatable conditions which can be measured. For this too, again an incubation is required as well as subsequently one or more washing steps in order to remove excess material. After such, an effected marking of the whole complex, the complex directly, or where appropriate, after an addition of a reagent, undergoes a measurement which gives insight on the type and quantity of the bound molecule, for example, the antibodies which are to be acquired.
The variation of measuring methods is likewise as large as the variation of method steps. Common to all, however, is a comparatively lengthy duration which is determined by the addition steps and the incubation. In practice, such methods require procedural times in the range of a few hours to several days. Due to the numerous different method steps and with this, the method parameters which are partly to be kept within small tolerances, such examinations as a rule can be only carried out by well qualified personnel.
Moreover, the quality of the results depends on the sum of the qualities of all the method steps. This causes the known methods to have a relatively high inaccuracy since the tolerances of the individual method steps acccumulate.
Yet another problem is that the test methods are designed such that the molecules bound on the solid phase are only provided for bonding to a molecule class, so that, for example, for recording various immune globulins various other methods must be carried out to bring about a good measurement. With regard to this the following literature is referred to:
R. H Burdon, P. H. van Knippenberg, eds.: Laboratory Techniques in Biochemistry and Molecular Biology, Vol.15; Elsevier Science Publishers B. V., Arnsterdam, New York, Oxford; 1988.
L. Hudson, F. C. Hay: Practical Immunology; Blackwell Scientific Publications, Oxford, London, Edinburgh, Melbourne, 1980.
Aside from the enormous time consumption, apparatus requirements and personnel intensive effort required by these known methods, there arises a considerable amount of waste (in particular, chemical or radioactive contaminated waste) which must be partly disposed of as special waste. The active substances, in fluid form, to be used with these methods are often unstable and must be protected from microbial infection. The energy consumption too is not inconsiderable, since the incubation steps as a rule must be carried out at body temperature, that is at 37 degrees Celsius and usually the apparatus must be kept at this temperature day and night. For a long time, one has been striving for a more simple and reliable test method.
A way which, at its beginning, seemed to hold out some promise was biosensory analysis. Biosensory analysis used semiconductors which determined a change of electrical characteristics of the molecules deposited on the substrate material as a basis for determining molecule bonding. However, this methodology has not brought the expected success because the changes of electrical characteristics occurring by way of selective molecule bonding are either not particularly significant or can only be measured inadequately. Therefore, in practice, such methods of bio-sensory analysis functioning on a semiconductor basis have only been carried out for two component systems in a limited manner. But these methods too are not very accurate. The semiconductor base required as the substrate has to be disposed of. What is needed, therefore, is an accurate and simple method and procedure for measuring binding reactions of atoms or molecules.
SUMMARY OF THE INVENTION
Against this background it is the object of the invention to provide a quantitative and/or qualitative determination of atoms and molecules with which, with comparatively little technical apparatus expenditure, this determination may be carried out simply and with a high reliability.
Accordingly, the invention provides for anchoring on a solid or fluid flat substrate, such molecules known per se it may also concern atoms with special applications, which are in the position to selectively bind other certain molecules or atoms these molecules are hereinafter described as receptor molecules. The substrate provided with these receptor molecules is then irradiated with electromagnetically polarized waves wherein the preferably reflected but also where appropriate the beamed through waves are received and acquired with regard to their state of polarization by way of a suitable device. Subsequently, the substrate with the receptor molecules anchored thereon are exposed to the material to be examined, for example, a blood sample. Afterwards, the substrate with the receptor molecules anchored thereon, as well as where appropriate the further molecules (ligands) or atoms bound in this way, are exposed to the electromagnetically polarized waves, whereafter ag
Meineke Dietmar
Riss Udo
Jacox Meckstroth & Jenkins
Swartz Rodney P.
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