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To directly radiolabel an anti-hepatoma mAb fragment HAb18 F(ab’)2 with 99mTc …


Biology Articles » Biophysics » Medical Biophysics » Direct technetium-99m labeling of anti-hepatoma monoclonal antibody fragment: a radioimmunoconjugate for hepatocellular carcinoma imaging

Abstract
- Direct technetium-99m labeling of anti-hepatoma monoclonal antibody fragment: a radioimmunoconjugate for hepatocellular carcinoma imaging

Direct technetium-99m labeling of anti-hepatoma monoclonal antibody fragment: a radioimmunoconjugate for hepatocellular carcinoma imaging

Hui Jie Bian1, Zhi Nan Chen1 and Jing Lan Deng2

1Cell Engineering Research Center, Basic Medical Department, Fourth Military Medical University, Xi’an 710032, Shaanxi Province, China
2Department of Clinical Nuclear Medicine, Xijing Hospital, Xi’an 710033, Shaanxi Province, China

World J Gastroenterol  2000;June6(3):348-352

Supported by National Natural Science Foundation of China, No.39700175

 

Subject headings: antibody, monoclonal; antibody fragments; technetium-99m; hepatocellular carcinoma; liver neoplasms; radioimmunoimaging

 

Abstract
AIM: To directly radiolabel an anti-hepatoma mAb fragment HAb18 F(ab’)2 with 99mTc by stannous-reduced method, and assess the stability, biodistribution and radioimmun oimaging (R).

METHODS: Immunoreactive fraction was determined according to Lin dmo’s method. Ellman’s reagent was used to determine the number of thiols in the reduced F(ab’)2. Labeling efficiency and homogeneity were measured by paper chromatography, sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) and autora diography. Challenge assay involved the incubation of aliquots of labeled antibo dy in ethylenediaminetetraacetate (EDTA) and L-cysteine (L-cys) solutions with different molar ratio at 37
for 1h, respectively. Investigations in vivo utilized nude mice bearing human hepatocellular carcinoma (HHCC) xenografts with gamma camera imaging and tissue biodistribution studies at regular intervals.

RESULTS: The labeling procedure was finished within 1.5h compared with the “pretinning” method which would take at least 21h. In vitro studies demonstrated that the radiolabeled mAb fragment was homogen eous and retained its immunoreactivity. Challenge studies indicated that 99mTc-labeled HAb18 F(ab’)2 in EDTA is more stable than in L-cys. Imaging and biodistribution showed a significant tumor uptake at 24h post-injection of 99mTc-labeled HAb18 F(ab’)2. The blood, kidney, liver and tumor uptakes at 24h were 0.56±0.09, 56.45±11.36, 1.43±0.27 and 6.57±3.01 (%ID/g)
respectively.

CONCLUSION: 99mTc-HAb18 F(ab’)2 conjugate prepare d by this direct method appears to be an effective way to detect hepatoma in nude mice model.
 

 

 


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