Chemistry: analytical and immunological testing – Involving an insoluble carrier for immobilizing immunochemicals – Carrier is organic
Reexamination Certificate
2000-04-25
2002-02-26
Venkat, Jyothsna (Department: 1627)
Chemistry: analytical and immunological testing
Involving an insoluble carrier for immobilizing immunochemicals
Carrier is organic
C435S025000, C435S189000, C210S649000, C210S650000, C210S651000, C210S652000, C210S653000, C210S654000, C210S655000, C210S500290, C210S500300, C210S500310, C210S500320
Reexamination Certificate
active
06350621
ABSTRACT:
FIELD OF THE INVENTION
The invention relates to (bio)chemical reagent solid phases which are comprised of a polymer or macromolecular NH
2
-containing support compound, on which coupling structures are bound via NH
2
groups covalently and which is further coupled with a receptor compound and/or indicator compound and thus the receptor and/indicator compounds can be coupled to the carrier. The invention also relates to a process for producing (bio)chemical reagent solid phases and their applications.
BACKGROUND OF THE INVENTION
With (bio)chemical reagent solid phases of the aforedescribed type, by the coupling of receptor compounds to the carrier compound, the receptors can be immobilized. Such immobilized receptor compounds have utility in a variety of fields and have been known for a long time. A typical field of use is in biotechnology, in (bio)chemical sensors and biochemical analysis as, for example, chromatography. Of interest are special immobilized biological compounds like enzymes or immunoproteins. With these, the (enzyme) catalytic function or complementarity is used for producing and/or purifying substances or by complementarity is used analytically for detection of analyte materials and for signal processes.
With respect to the immobilization process and the composition of reagent solid phases, there is an enormous variation described in the technical literature. They are produced above all by the modification:
of the structure of the carrier compound as, for example,
macromolecular carriers of an organic or inorganic nature like collagens, dextrans or porous glass and the like and
polymeric carriers of polar and nonpolar nature like polyamides, polyurethanes or polystyrenes, polyethylenes or the like;
of an immobilizing kind or utilizing an immobilization process as for example
immersion or adsorption
electrostatic or covalent fixation, for example, via so-called bifunctional coupling compounds typically by means of glutaricdialdehyde or the like or
via the use of conventional synthesis reaction, e.g. substitution reactions, diazotizing reactions or the like;
of the receptor compound, for example,
biocompounds like enzymes (oxide reductases, proteases or the like or immunoproteins (antibodies) or
organic receptor compounds like crown ethers for ions or the like.
See Hermanson, G. T. et al (Ed.): Immobilized Affinity Ligand Technics, Academic Press, 1992 or Chibatu, I. (Ed.): Immobilized Enzymes, Research and Development, Kodanska Scientific Books, 1978).
Especially for the immobilization of biocompounds like enzymes or immunoproteins, polysaccharides, especially cellulose can be used as the carrier compound. For all of the important immobilization categories like
carrier inclusion,
ion association
covalent bonding
a multiplicity of variations have been found also with respect to the biocompounds which are incorporated. Table 1, assembles a collection of selected possibilities (compare the above-cited Hermanson, G. T. et al).
TABLE 1
Example of Enzyzme-Immobilizations on Cellulose
Derivatives
Cellulose Derivative
Immobilization type
Enzyme Example
Butylacetatcellulose
Carrier inclusion
Urease, etc.
Nitrocellulose
Carrier inclusion
Lactase,
Asparaginase, etc.
Collodium
Carrier inclusion
Lactase, Urease etc
DEAE-Cellulose
Ion-inclusion
Catalase, Invertase,
Pepsin, etc.
TEAE-Cellulose
Ion inclusion
Aspartase, etc.
Covalent Bonding To:
p-Aminobenzylcellulose
Diazo-Reaction
Catalase, Invertase,
Pepsin, Aspartase
etc
p-Aminobenzoylcellulose
Diazo-Reaction
Trypsin,
Chymotrypsin, etc.
Bromcyan-activated
Bromcyan activation
Xanthin-Oxidase,
Cellulose
Lactase, Dextranase
etc.
CM-Cellulose,
Carbodiimide method
Peroxidase, etc.
AE-Cellulose
Nitrocellulose-Membranes
Membrane inclusion
Glucose-Oxidase
Peroxidase, etc
Bromcyan-activated
Meuibrane inclusion
Lactase, Trypsin etc
Cellulose Membranes
Bromcyan activation
The columns in Table 1 can be expanded with respect to further cellulose derivatives, for example, cellulose carbonate chloracetylcellulose, bromacetylcellulose, etc., and also with reference to further biocompounds, for example, further enzyme species, NAD- and pyridoxal-phosphate coenzyme, vitamin B
12
, immunoproteins and the like.
In the forefront of the examples given, are preparative or analytic, e.g. chromatographic, applications in which the carrier compounds with receptor compounds or enzymes will depend only on the loading density and without special molecular geometric considerations.
With respect to the functional requirements, for example with the enzyme immobilization process, very different scales apply depending upon the field of use. Target criteria of an effective enzyme immobilization are
a precise folding of the proteins,
a free substrate accessibility of the active center,
an effective product recovery,
carrier fixation on the enzyme-molecular periphery,
a high enzyme loading density per unit of carrier matrix surface and
in the case of use as a sensor, a carrier matrix with signal structure features which affords a maximum optical or electronic transfer signal transduction.
The use of reagent solid phases in (bio) chemical sensor technology requires that, for each sensor development for a respective analyte, a new structure optimization based upon the aforestated criteria; the sensor solution for one analyte then can hardly be used for other analytes without further translation. This drawback in the state of development in the (bio) chemical sensor field makes the use of many scientifically determined sensor developments limited in practice, because there is a significant gap between the requirements of the sensor user and the generally limited functional stability and cross sensitivity of the sensors.
A solution of this problem was expected from sensor transducers of a “measurement-tailored” supermolecular structure with improved signal transmission characteristics. Such a transducer is known, for example, from enzyme electrode developments, using polymeric carrier compounds with so-called electron mediator structures, e.g. ferrocene derivatives, and immobilized enzymes, for improved signal transduction by reduction of the electron transition “barriers”, and the measurement electrode (see B. Frew, J. E. and Hill, H. A. O. (1987); Electrochemical Biosensors, Anal. Chem. 59, 933A; Frew, J. E. and Hill, H. A. O. (1987): Electron-transfer-Biosensors, Phil. Trans. R. Soc. B316, 95; Bockris, J. O'M. and Khan, S. U. M. (Ed.): Surface Electro-chemistry—A Molecular Level Approach, Plenum Press, 1993).
Significantly more complex supermolecular structures are required in the case of (bio) chemical glass fiber sensor transducers. The higher complexity is based upon the fact that an enzyme protein which possesses the requisite function and in most cases in addition, a further structure component, for example, an indicator structure, must be provided in a well-defined molecularly geometric positioning of the structures relative to one another on the polymer or macromolecular carrier matrix. Exceptions in which an additional signal structure is not required form analyte recognition structures wherein the analyte recognition and the optical signal transfer are inherent in the structure; for these up to now there have however been few examples.
In the field of glass fiber sensor technology there have already been reagent solid phases used which have cellulose derivatives as carrier compounds, on which pH indicators have been immobilized via vinyl sulfonyl groups as couplers (Weigl, B. H. et al. (1993): Robust Carbon Dioxidoptrode Based on a Covalently Immobilized pH-Indicator, SPIE-Vol. 2068, 2). In addition, the use of cellulose acetate membranes in combination with pH indicators has been described (Sansubrino, A. and Mascini, M. (1994): Development of an Optical Fiber Sensor for Ammonia, Urea, Urease and IgG, Biosens. Bioelectron. 9, 207).
The development of (bio) chemical glass fiber sensors is of special interest because of their capacity for integration, the possibility of a high degree of miniaturization and further advantageous characteristics, the
Berlin Peter
Klemm Dieter
Tiller Jörg
Dubno Herbert
Forshungszentrum Mulich GmbH
Garcia Maurie E.
Myers Jonathan
Venkat Jyothsna
LandOfFree
(Bio)chemical reagent solid phases, process for their... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with (Bio)chemical reagent solid phases, process for their..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and (Bio)chemical reagent solid phases, process for their... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2940883