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G L Griffiths

Publications and source records attributed to G L Griffiths.

At least 37 records · Page 2Linked to original sources
PubMed1994

Technetium-99m, rhenium-186, and rhenium-188 direct-labeled antibodies.

BACKGROUND: Antibody sulfhydryl groups can act as effective carriers of reduced technetium and rhenium species for radioimmunodetection and radioimmunotherapy. METHODS: Intact immunoglobulin G and fragments were labeled with the isotopes and examined in vitro and in vivo. RESULTS: Technetium bound to intact immunoglobulin G was found to be the most stable species in vitro, but in vivo, clearances of technetium and rhenium bound to intact antibody were similar. Serum clearances were faster than those seen for the corresponding radioiodinated antibodies. In vivo clearance rates of the radiolabeled fragments were similar, with kidney uptake and retention seen. Rhenium-labeled antibodies, despite a greater tendency toward in vitro reoxidation than technetium-labeled antibodies, did not show enhanced kidney clearance in animal models. Rhenium-188 and technetium-99m were obtained from similar generator systems in carrier-free form. Using rhenium-188 spiked with cold rhenium, it was determined that approximately one rhenium atom per molecule of antibody can be conjugated directly. Rhenium-186 also was coupled at almost a 1:1 ratio to antibody. CONCLUSIONS: Only radiolysis concerns will limit the amount of rhenium-188 conjugated to antibody. Large doses of antibody will be necessary to deliver rhenium-186 at this isotope's currently available specific activity. Otherwise, higher specific activity rhenium-186, and/or greater loading capacity of rhenium-186 onto antibody, will be needed to generate the type of product that will be usable at a clinical dose of several hundred millicuries.

Animals↗
PubMed1994

Processing of antibody-radioisotope conjugates after binding to the surface of tumor cells.

BACKGROUND: Previous experiments indicated that most antibodies binding to cell surface antigens are internalized gradually and degraded within lysosomes, with a half-life of degradation of approximately 1 day, for most antibodies. The research discussed in this article extended our studies to eight additional antibodies reacting with six different antigens, including three antigens anchored in the membrane by glycosyl-phosphatidylinositol. The authors also tested antibodies labeled with 111indium, as well as 125iodine, to determine whether different radiolabels would be processed differently. METHODS: Antibodies were radiolabeled with 125I or with 111In bound to benzyl-DTPA. After binding to the surface of tumor cells in vitro, excess antibody was washed away, and the fate of the radiolabel was investigated over periods of 3-7 days. Radiolabel released into the supernatant or retained by the cells was analyzed to determine whether it was still on intact antibody. RESULTS: In 13 of the 15 antibodies that were tested, a similar pattern of irreversible binding and gradual catabolism was observed. Iodine conjugated to antibodies was released rapidly from the cell after antibody catabolism. In contrast, the 111In was retained within the cell much longer than 125I, with the rate of degradation and release into the medium being at least fivefold slower. More than 50% of the bound 111In was still present on the cells after 7 days. Biochemical analysis of the retained 111In extracted cells after 4-6 days demonstrated that it was no longer associated with antibodies and was in a low molecular weight form, probably still associated with the chelator benzyl-DTPA. CONCLUSIONS: Different radiolabels are processed by tumor cells differently, after catabolism of the antibody to which they originally were conjugated. The data suggest that the prolonged retention of 111In, relative to that of 125I, is due not to deiodination of iodine conjugates, but rather to intracellular retention of catabolic products containing 111In, perhaps within lysosomes. The use of radioisotopes that are retained within cells after antibody internalization and degradation may improve both radioimmunodetection and radioimmunotherapy of cancer.

Antibodies, Monoclonal↗
PubMed1994

Preparation of a pure 99mTc-F(ab')2 radioimmunoconjugate by direct labeling methods.

Intact IgG and Fab' can be labeled directly with 99mTc to give quantitative incorporation of radioactivity into the protein. With F(ab')2 the reductive conditions yield a mixture of 99mTc-F(ab')2 and 99mTc-Fab'. We now report a direct labeling method to produce only 99mTc-F(ab')2 in quantitative yield and contaminated with 99mTc-Fab'. The properties, stability and biodistribution of the 99mTc-F(ab')2 have been compared to 99mTc-Fab'. This new technology will allow us to compare technetium direct-labeled IgG, F(ab')2 and Fab' derivatives of the same antibody for radioimmunodetection.

Animals↗
PubMed1994

The processing and fate of antibodies and their radiolabels bound to the surface of tumor cells in vitro: a comparison of nine radiolabels.

UNLABELLED: Processing radiolabeled degradation products is the key factor affecting retention of antibodies within the cell. In this study, we have analyzed the processing of antibodies labeled in nine different ways. METHODS: Antibodies were labeled with three different radioisotopes and seven different forms of 125I. Eight of the radiolabels (except 188Re) were conjugated to the same antibody, MA103, and tested on the renal carcinoma cell line SK-RC-18 and/or the ovarian carcinoma cell line SK-OV-6. Rhenium conjugation utilized the antibody RS7, the target cell line ME180 and three of the other radiolabels were also tested with this antibody-target cell combination for comparison. RESULTS: Iodine conjugated to antibodies by conventional methods was rapidly released from the cell after antibody catabolism. In contrast, iodinated moieties, such as dilactitol-tyramine and inulin-tyramine were retained within cells four to five times longer. CONCLUSIONS: The use of radiolabels that are trapped within cells after antibody catabolism can potentially increase the dose of radiation delivered to the tumor, from the same amount of radioactivity deposited by a factor of four or five. The prolonged retention of 111In relative to 125I is not due to deiodination of iodine conjugates, but rather to intracellular retention of catabolic products containing 111In, perhaps within lysosomes.

Acetylgalactosamine↗
PubMed1991

Direct radiolabeling of monoclonal antibodies with generator-produced rhenium-188 for radioimmunotherapy: labeling and animal biodistribution studies.

The use of 188Re from an alumina-based 188W/188Re generator has been investigated for antibody radiolabeling. It was found that, with simple labeling techniques, 188Re can be used immediately after elution. The direct radiolabeling of intact antibodies with 188Re is described. Lyophilized antibody preparations have been reconstituted with 188Re taken directly from the generator at specific activities of up to 15 mCi of 188Re per mg of antibody. Radiolabeling yields of 90 to 98% have been obtained, with the incorporation rate being dependent upon time and the relative concentrations of the reagents. It was determined that the conjugates were immunoreactive and stable when challenged by serum in vitro, with 188Re-immunoglobulin G showing adequate resistance to reoxidation with no transfer of 188Re to serum protein. 188Re-antibody conjugates were shown to clear from the blood faster than the corresponding 131I-labeled antibody, giving rise to good tumor/nontumor ratios at 24 to 72 h postinjection, while serum samples taken from the animals have shown that the circulating 188Re remained bound to immunoglobulin G. The combination of the technologies of the 188W/188Re generator, the direct labeling methodology, and the use of single-vial lyophilized antibody makes the use of 188Re-radiolabeled monoclonal antibodies a simple and convenient method of cancer radioimmunotherapy with a beta-emitting radionuclide.

Animals↗
PubMed1990

Selection of a DTPA chelate conjugate for monoclonal antibody targeting to a human colonic tumor in nude mice.

Our previous studies with a 90Y-labelled antibody against carcinoembryonic antigen (CEA) conjugated to the cyclic anhydride-DTPA (CA-DTPA) indicated that the accretion of 90Y in the bone may limit the application of 90Y-labelled antibodies for therapy. In this report, we have compared the tumor targeting of CA-DTPA-conjugated antibody to antibody conjugated with 4 isothiocyanatobenzyl (ITC-Bz) derivatives of DTPA in nude mice bearing a human colonic tumor xenograft. In biodistribution studies using an 111In-labelled anti-CEA murine monoclonal antibody (MAb), the CA-DTPA-conjugated MAb showed lower tumor uptake, faster blood clearance, and higher accretion in the liver than any of the 4 ITC-Bz-DTPA-conjugated MAbs. There were smaller differences among the 4 ITC-Bz-DTPA conjugates. Whole-body autoradiography of animals given 90Y-MAb prepared with the CA-DTPA or the ITC-Bz-DTPA showed less radioactivity in the bone with the ITC-Bz-DTPA-MAb than the CA-DTPA-MAb. 90Y uptake in the bone corresponded with regions of low proliferative activity as defined by 3H-labelled thymidine, suggesting that the 90Y was in the cortex rather than the marrow. These studies clearly show an advantage of the ITC-Bz-DTPA derivatives for 90Y and 111In labelling of MAbs.

Animals↗
PubMed1989

Salinomycin poisoning in point-of-lay turkeys.

Salinomycin poisoning occurred in a flock of 700 point-of-lay turkeys; 400 birds died over 7 days as a result of consuming feed contaminated with 50 ppm salinomycin. No gross lesions were detected. Histologic evidence of a myopathy was most readily detected in leg muscles of turkeys 5 to 7 d after ingesting salinomycin. Feeding trials were undertaken and individual susceptibility to the drug was found to vary greatly. In affected birds the plasma concentrations of creatine kinase (CK) and aspartate aminotransferase (AST) were found to be in the range of 500,000 to 2,500,000 IU/l and 9000 to 25,000 IU/l, respectively. The marked increase in the plasma activities of these enzymes preceded histological evidence of segmental muscle necrosis.

Animal Feed↗
PubMed1985

Blood content of broiler chicken carcases prepared by different slaughter methods.

Two trials using broiler chickens were undertaken to determine if variations in commercial slaughter techniques affected either the appearance or the residual blood content of the carcases produced. In the first trial groups of broilers were either stunned (55, 80 or 105 volts), electrocuted (240 volts) or anaesthetised and the subsequent bleed-out volumes after one, two, three and four minutes exsanguination were measured. The method of slaughter made no difference to the amount of blood lost after venesection. Haemoglobin content of the pectoral muscle was used to measure residual blood and was uniformly low in all groups (0.7 to 0.16 mg haemoglobin/g of muscle). In the second trial groups of chickens were either stunned and bled out, electrocuted, electrocuted and bled out, decapitated or halal killed before processing. The edible portions of the carcases were minced and the average haemoglobin content for each group was 0.19, 0.36, 0.22, 0.17 and 0.17 mg/g of soft tissue. The appearance of chickens that were stunned and bled out was the same as that of chickens that were electrocuted and bled out. The authors conclude that killing chickens by electrocution before exsanguination does not adversely affect the carcase appearance. Furthermore, electrocution ensures that broiler chickens are humanely slaughtered.

Abattoirs↗
PubMed1984

Vibriobactin, a siderophore from Vibrio cholerae.

A novel siderophore (microbial iron transport compound) has been isolated from low iron cultures of Vibrio cholerae. Belonging to the catecholamide family of chelators, it has been shown to contain three residues of 2,3-dihydroxybenzoic acid and two residues of threonine. Both threonine moieties are present in the form of oxazoline rings. Furthermore, the polyamine backbone of the molecule was proved to be not spermidine, but the rare N-(3-aminopropyl)-1,3-diaminopropane, norspermidine. The structure of the new siderophore has been determined to be N-[3-(2,3-dihydroxybenzamido)propyl]-1, 3-bis[2,3-dihydroxyphenyl)-trans-5-methyl-2-oxazoline-4-carboxamido]prop ane. The compound has been given the trivial name vibriobactin. Mutants defective in the synthesis and utilization of vibriobactin were isolated. In an iron-limited environment V. cholerae was found to respond more strongly to vibriobactin, agrobactin, and ferrichrome than to enterobactin.

Catechols↗
PubMed1984

A survey of slaughter procedures used in chicken processing plants.

Information was collected on the slaughter procedures used in 12 processing plants in Australia. The processing line speed varied from 25 to 125 birds/minute; the voltage in the electrified waterbath varied from 55 to 165 V; the duration of stunning was 2.0 to 7.5 seconds and the time allowed for exsanguination was 45 to 180 seconds. As a result of the variables, some birds were dead on leaving the stunner and prior to exsanguination, while it was probable that others were not dead before entering the scald tank.

Abattoirs↗