Biomedical subjects

P Pivarnik

Publications and source records attributed to P Pivarnik.

5 recordsLinked to original sources

PubMed2000

Evaluation of antibody immobilization methods for piezoelectric biosensor application.

The immobilization of anti-Salmonella antibodies by two methods were studied and evaluated for their potential use in a piezoelectric biosensor. The optimum temperature-time combinations for the highest immobilization yields were determined for both methods. Protein A binding was found to be 67.4+/-3.8% on the gold surface which then allowed an immobilization of 42.1+/-2.09% antibody. The degree of antibody immobilization via surface aldehyde groups of glutaraldehyde (GA) on a precoated quartz crystal with polyethylenimine (PEI) was 31.6+/-0.3%. A piezoelectric probe was designed and used in dry assays to observe the frequency change due to addition of mass by the immobilization layers. The frequency changes recorded showed a better reproducibility and less added mass for the Protein A method. The frequency decrease due to microg of added antibodies was compared to frequency decrease calculated by the Sauerbrey equation. The experimental data was found to be only approximately 8% of theoretical data. The functionality of the immobilized antibodies with the Protein A method was tested with S. typhimurium in a wet chamber and the frequency decrease was compared to results of a similar system activated with PEI-GA immobilization. The frequency decreases with S. typhimurium concentration of approximately 1.5 x 10(9) CFU/ml were 50+/-2 Hz and 44+/-3 Hz for the Protein A method and PEI-GA method, respectively. It was concluded that although both methods resulted in comparable activities in terms of % immobilized protein and frequency decreases due to Salmonella binding, the Protein A method was favorable due to stability and better reproducibility of the immobilization layers.

Antibodies, Bacterial↗

PubMed1999

Colloidal gold filtrates as metal substrates for surface-enhanced infrared absorption spectroscopy.

A new method for obtaining surface-enhanced infrared absorption (SEIRA) spectra of antibodies and antibody/antigen complexes has been developed. Antibodies attached to colloidal gold particles and then collected by filtration onto porous polyethylene membranes show enhanced spectral bands at 1080 and 990 cm-1 regardless of the antibody specificity. Attachment of a model antigen, glucose oxidase, to its specific antibody/colloid complex prior to collection produces enhanced bands at 1540, 1395, and 1250 cm-1. Similarly, when the antigen Salmonella is attached to its specific antibody/colloid complex prior to collection, a new enhanced band is observed at 1015 cm-1. Similarities and differences of the SEIRA spectra obtained on gold colloid are compared to previous work on gold films.

Antibodies↗

PubMed1998

Immunoassays based on surface-enhanced infrared absorption spectroscopy.

A new type of biosensor for pathogens has been developed. The sensor produces spectral fingerprints of biological systems by using surface-enhanced infrared absorption (SEIRA) spectroscopy. Antibodies were immobilized onto a 10-nm-thick film of gold which had been previously deposited on a Si wafer. SEIRA spectra of the antibodies measured in the external reflection mode exhibited two new bands at 1085 and 990 cm-1. These new bands were observed with p-polarized radiation but were absent with s-polarized radiation. The spectrum of water on the surface of the sensor was observed under both directions of polarization. The sensor was first tested with a model system consisting of glucose oxidase (GOX) and the antibodies for glucose oxidase (anti-GOX). In addition to the bands due to the anti-GOX at 1085 and 990 cm-1, new bands were observed at 1397, 1275, and 930 cm-1 when the GOX antigens were present. The same type of sensor was prepared for Salmonella (SAL) by immobilizing antibodies for Salmonella (anti-SAL) on a gold-surfaced Si water. The SEIRA spectra for anti-SAL antibodies were very similar to those for anti-GOX, with bands at 1085 and 990 cm-1; however, a sharp new band was observed at 1045 cm-1 after the sensor was exposed to the SAL antigens. In addition to specific new bands due to antigens, both GOX and SAL sensors exhibited changes in the regions of water absorptions at approximately 3500 and 850 cm-1 when the antigens were present.

Animals↗

PubMed1998

Acoustic standing-wave enhancement of a fiber-optic Salmonella biosensor.

An enhanced fluorescent fiber-optic biosensor system using ultrasonic concentration of particles and cells has been developed and applied in the detection of Salmonella typhimurium. A biosensor test chamber also serves as an ultrasonic standing-wave cell that allows microspheres or cells to be concentrated in parallel layers or in a column along the axis of the cell. A fiber probe along the axis delivers laser excitation to fluorescent-labeled antibodies of Salmonella and collects the fluorescent signal. The labeled-antibodies themselves do not respond to the ultrasound, but, when attached to Salmonella cells, the Salmonella-antibody complexes can be moved acoustically to the axis of the cell, increasing the fluorescent signal. In a second, more robust, type of immunoassay, the Salmonella-labeled-antibody complexes attach to unlabeled antibodies that have been immobilized on the surface of polystyrene microspheres. This entire structure can be manipulated acoustically and the increase in the fluorescent signal, which can be an order of magnitude, indicates the presence of Salmonella.

Biosensing Techniques↗

PubMed1994

Pharmacodynamics of piperacillin alone and in combination with tazobactam against piperacillin-resistant and -susceptible organisms in an in vitro model of infection.

The pharmacodynamics of dosage regimens of piperacillin alone or in combination with tazobactam against piperacillin-resistant or -susceptible bacteria were studied in an in vitro model of infection. Experiments were conducted by using a fixed daily exposure of 12 g of piperacillin, given as 3 g alone or in combination with tazobactam at 0.375 g every 6 h, or the same total dose of the combination given as 4 g of piperacillin plus 0.5 g of tazobactam every 8 h. The addition of tazobactam to piperacillin, irrespective of the dosing interval, did not alter the killing of piperacillin-susceptible organisms (Escherichia coli J53 and Pseudomonas aeruginosa ATCC 27853). In contrast, experiments with an isogenic TEM-3-containing transconjugant of E. coli J53 (E. coli J53.2-TEM-3) that was resistant to piperacillin (MIC, 128 micrograms/ml) showed that the addition of tazobactam resulted in bacterial killing similar to that observed with the wild-type strain. Although tazobactam concentrations fell to less than 4 mg/liter (the concentration associated with a reduction in the piperacillin MIC from 128 to 2 mg/liter) 2 to 3 h after a dose, a similar degree of bacterial killing was observed when the same total 24-h dose of piperacillin-tazobactam was fractionated into dosing intervals of every 6 or 8 h. Investigations with Staphylococcus aureus 7176 (piperacillin MIC, 128 micrograms/ml) showed that the addition of tazobactam, again irrespective of dosing interval, also resulted in net bacterial killing which was not seen with piperacillin alone. These data support the use of extended dosing intervals (every 8 h) of piperacillin-tazobactam in the treatment of infections caused by piperacillin-resistant bacteria.

Bacteria↗