99mTc Direct radiolabeling of PR81, a new anti-MUC1 monoclonal antibody for radioimmunoscintigraphy
Authors
Abstract:
Introduction: Monoclonal antibodies labeled mainly with 99mTc are being widely used as imaging agents in nuclear medicine. Recently PR81 was introduced as a new murine anti-MUC1 MAb against human breast carcinoma. This antibody reacts with the tandem repeat of 20-mer peptide of protein backbone, and is of IgG1 class, with an affinity of 2.19×10-8 M-1. Due to high specific reactivity, this antibody was shown to have high diagnostic potential in nuclear medicine. As the first step towards the use of an antibody for imaging purposes it has to be labeled with a radioisotope. In this study we have investigated a method for optimum labeling of this MAb with 99mTc. Materials and Methods: The Ab reduction was performed with 2-mercaptoethanol (2-ME) at a molar ratio of 2000:1 (2-ME:MAb) and reduced Ab was labeled with 99mTc via stannous tartrate as a transchelator. The labeling efficiency was determined by ITLC. The integrity of reduced MAb was checked by means of SDS-PAGE and gel filtration chromatography (FPLC) on Superose 12 HR 10/30 (purity>99%). The amount of radiocolloids were measured by cellulose nitrate electrophoresis. In vitro stability of labeled product was checked in time intervals over 24 hrs by ITLC. Radioimmunoassay was used to test immunoreactivity of the labeled MAb. Biodistribution of the radiolabeled MAb was studied in normal mice at 4 and 24 hrs post-injection. Results: The labeling efficiency was %96±2.8 with high invitro stability (%82±2.7 in 24 hrs) and less than %2 radiocolloids were found. There was no Ab fragmentation due to reduction procedure. Both labeled and unlabeled MAbs were able to compete for binding to the MUC1. Biodistribution studies in normal mice showed that there was no significant accumulation in any organ. Conclusion: Because of no significant loss of immunoreactivity of MAb due to labeling procedure, high in vitro stability and no accumulation in vital organs, 99m-Tc-PR81 may be a promising candidate for radioimmunoscintigraphy studies of breast cancer.
similar resources
99mtc direct radiolabeling of pr81, a new anti-muc1 monoclonal antibody for radioimmunoscintigraphy
introduction: monoclonal antibodies labeled mainly with 99mtc are being widely used as imaging agents in nuclear medicine. recently pr81 was introduced as a new murine anti-muc1 mab against human breast carcinoma. this antibody reacts with the tandem repeat of 20-mer peptide of protein backbone, and is of igg1 class, with an affinity of 2.19×10-8 m-1. due to high specific reactivity, this antib...
full textRadioimmunoscintigraphy of Breast Tumor Xenografts in Mouse Model by 99mTc Direct Radiolabeling of a Monoclonal Antibody PR81
Introduction: The radioimmunoscintigraphy (RIS) has found widespread clinical applications in tumor diagnosis. Human epithelial mucin, MUC1, is commonly over expressed in adenocarcinoma including 80% of breast cancers and represents a useful target for RIS. The PR81 is a new murine anti-MUC1 monoclonal antibody that was found to react with the membrane extracts of se...
full textradioimmunoscintigraphy of breast tumor xenografts in mouse model by 99mtc direct radiolabeling of a monoclonal antibody pr81
introduction: the radioimmunoscintigraphy (ris) has found widespread clinical applications in tumor diagnosis. human epithelial mucin, muc1, is commonly over expressed in adenocarcinoma including 80% of breast cancers and represents a useful target for ris. the pr81 is a new murine anti-muc1 monoclonal antibody that was found to react with the membrane extracts of se...
full textتولید و کنترل کیفی آنتی بادی مونوکلونال نشان دار با تکنسیم 99m علیه سلول های سرطانی کولون
Background and purpose: Binding a monoclonal antibody to tumor associated antigens is an effective method for cancer therapy because these agents can specifically target malignant cells. In fact, monoclonal antibodies are effective agents for diagnosis, grading and treatment of different kinds of cancers. In this research, a new monoclonal antibody against colon cancer cells was prepared and ...
full textEpithelial mucin-1 (MUC1) expression and MA5 anti-MUC1 monoclonal antibody targeting in multiple myeloma.
Multiple myeloma (MM) is the second most common hematological cancer in the United States. It is typically incurable, even with myeloablative chemotherapy and stem-cell transplantation. The epithelial mucin-1 (MUC1) glycoprotein is expressed by normal and malignant epithelial cells but has also been shown to be expressed by MM cells. MUC1 is a useful antigenic target in solid tumors for clinica...
full textMy Resources
Journal title
volume 13 issue 1
pages 7- 16
publication date 2005-01-01
By following a journal you will be notified via email when a new issue of this journal is published.
Hosted on Doprax cloud platform doprax.com
copyright © 2015-2023