Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Blood proteins or globulins – e.g. – proteoglycans – platelet...
Reexamination Certificate
1995-07-25
2004-06-15
Ponnaluri, Padmashri (Department: 1639)
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
Blood proteins or globulins, e.g., proteoglycans, platelet...
C530S389800, C435S007100, C435S007900, C435S007920, C436S518000, C436S536000, C436S547000, C436S548000
Reexamination Certificate
active
06750328
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
The invention relates generally to antibody assays, and more particularly, to monoclonal antibodies and antibody assays for the detection of cationic, anionic and nonionic water treatment polymers.
2. Description of the Prior Art
As described in U.S. Pat. Nos. 4,756,881 and 4,752,443, water-soluble sulfonated copolymers of acrylic acid and acrylamide (hereinafter referred to as “sulfonated copolymers”) are used in the treatment of industrial cooling water to prevent corrosion and mineral deposits (scale). Generally, the active sulfonated copolymers remove dissolved minerals from the cooling water by complexing with the mineral. Over time, the complexation sites of the sulfonated copolymer molecules become saturated and the copolymer molecules become inactive, unable to remove any additional minerals from the cooling water.
To prevent corrosion and scale damage to machinery, as the polymers are inactivated they must be removed and replaced by active sulfonated copolymer. Thus, active sulfonated copolymer must be continually fed into the cooling water to replace the inactive sulfonated copolymer. Maintaining the proper feed level for the active sulfonated copolymer is essential for optimum performance of the cooling water system. An improper feed rate can lead to serious problems. For example, insufficient active sulfonated copolymer can result in the water treatment being overwhelmed by dissolved minerals, thereby causing severe corrosion or scale deposit. On the other hand, maintaining too high a level of the active polymer is very expensive and is an inefficient method for treating industrial cooling water.
Although several methods are available for determining the total concentration of sulfonated copolymer in an industrial cooling water system, i.e., active plus inactive sulfonated copolymer, these techniques are unsatisfactory since they only determine the concentration of total sulfonated copolymer, and do not measure the concentration of the active sulfonated copolymer. Moreover, these methods suffer from lack of specificity or poor sensitivity. For example, the older methods for detecting sulfonated copolymers include colloid titration with PVSK, complexation with Hyamine 1622, or reaction of excess magnesium with chrome azurol S. The above tests detect any polyanionic material and have a detection threshold of about 50 ppm polymer. Presently, the total amount of active sulfonated polymer in an industrial cooling water system cannot be inexpensively and rapidly determined.
Cationic polymers are also useful in many areas of industrial water treatment. These areas include paper manufacture, effluent stream clarification, sludge dewatering, mineral process and many others. Excessive amounts of cationic polymers may cause problems in waters discharged to the environment. It is therefore desirable to know with specificity and precision the amount of residual cationic polymer in a sample. Many prior art methods of determining cationic polymer concentrations in waste water and other water treatment systems suffer from lack of specificity or poor sensitivity as with the sulfonated copolymers described above. Desirably, a method that would solve the problems of sensitivity and specificity described for all types of polymer would be available.
SUMMARY OF THE INVENTION
One aspect of the present invention is directed to a monoclonal antibody having an affinity to water treatment polymers. These polymers may be sulfonated copolymers of acrylamide and acrylic acid or any generally available water treatment polymer. The monoclonal antibodies of the present invention have an absolute specificity for the water treatment polymer and a lower limit of detection in the femtogram per ml range when used in some detection formats.
The monoclonal antibodies of the invention are produced by hybridoma cell lines. One preferred hybridoma cell line of the invention is hybridoma cell line 6E2-H1-G4. Another preferred hybridoma cell line of the invention is hybridoma cell line 6D12-H9-H3. Still other preferred cell lines include M11.2 and D8.2.
Another aspect of the invention is directed to a method of manufacturing a monoclonal or polyclonal antibody having an affinity to a water treatment polymer. The inventive method includes the steps of: a) immunizing a mammal with a substance that will elicit an antibody response to a water treatment polymer attached to a carrier protein; b) preparing a hybridoma cell producing the monoclonal antibody from cells removed from the immunized mammal; c) cloning the hybridoma cell to produce a hybridoma cell line; and d) extracting the monoclonal antibody from the hybridoma cell line. According to one preferred embodiment of the invention, the mammal is a mouse.
A further aspect of the invention is directed to a process for the determination of the presence or concentration of a water treatment polymer in a fluid. The inventive process includes the step of incubating a sample of the fluid containing the water treatment polymer with a monoclonal or polyclonal antibody having an affinity for the water treatment polymer.
REFERENCES:
patent: 4752443 (1988-06-01), Hoots et al.
patent: 4756881 (1988-07-01), Hoots et al.
patent: 5593850 (1997-01-01), Wetegrove et al.
patent: 6146903 (2000-11-01), Weatherbury et al.
patent: 6420530 (2002-07-01), Weatherby et al.
patent: 0559249 (1992-02-01), None
Balakrishnan Krishna
Carlin Robert J.
Stave James W.
Wetegrove Robert L.
Wu Ming-Hsien
Kilpatrick & Stockton LLP
Ponnaluri Padmashri
Strategic Diagnostics Inc.
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