The Relationship of Glycosylation and Isoelectric Point with Tumor Accumulation of Avidin (original) (raw)
Avidin Targeting of Intraperitoneal Tumor Xenografts
2000
Background: Lectins (proteins that bind specific sugar molecules on glycoproteins and glycolipids) are expressed at various levels on the surface of tumor cells. Conjugation of cytotoxic agents to glycoproteins recognized by lectins could be useful in the treatment of tumors. Avidin (a highly glycosylated, positively charged protein found in egg white) contains terminal N-acetylglucosamine and mannose residues that bind to some lectins. In this study, we tested the ability of avidin, labeled through conjugation to radioactive biotin (a B vitamin), to target intraperitoneal tumors. Methods: Biotin was radioactively labeled with 111 In. Four tumor models (one ovarian, one lung, and two colon) were established in nude mice by intraperitoneal injection of cultured cancer cells. The following two approaches were used in the intraperitoneal administration of avidin: 1) radioactive biotin-avidin conjugates were injected and 2) avidin was injected 1-24 hours before the injection of radioactive biotin (avidin pretargeting; avidin-biotin conjugates formed in vivo). The distribution of injected radioactivity in the tissues of treated animals was assessed. Results: Radiolabeled avidin localized highly and rapidly in the tumors. More than 50% of the administered dose of avidin-biotin conjugate accumulated per gram of tumor tissue 2 hours after injection; high tumor uptake of radioactivity was observed up to 24 hours after conjugate injection. In contrast, accumulation of radioactivity in normal tissues was low, yielding high tumor to nontumor ratios. With avidin pretargeting, accumulation of radioactivity in the liver, kidney, and spleen was reduced to a greater extent than that in the tumor, and tumor to nontumor ratios were increased. Conclusions: Avidin may be a promising vehicle for the delivery of radioisotopes, drugs, toxins, or therapeutic genes to intraperitoneal tumors. [J Natl Cancer Inst 1998;90:25-29]
Chemical Linkage to Injected Tissues Is a Distinctive Property of Oxidized Avidin
PLOS One, 2011
We recently reported that the oxidized avidin, named AvidinOXH, resides for weeks within injected tissues as a consequence of the formation of Schiff's bases between its aldehyde groups and tissue protein amino groups. We also showed, in a mouse pre-clinical model, the usefulness of AvidinOX for the delivery of radiolabeled biotin to inoperable tumors. Taking into account that AvidinOX is the first oxidized glycoprotein known to chemically link to injected tissues, we tested in the mouse a panel of additional oxidized glycoproteins, with the aim of investigating the phenomenon. We produced oxidized ovalbumin and mannosylated streptavidin which share with avidin glycosylation pattern and tetrameric structure, respectively and found that neither of them linked significantly to cells in vitro nor to injected tissues in vivo, despite the presence of functional aldehyde groups. The study, extended to additional oxidized glycoproteins, showed that the in vivo chemical conjugation is a distinctive property of the oxidized avidin. Relevance of the high cationic charge of avidin into the stable linkage of AvidinOX to tissues is demonstrated as the oxidized acetylated avidin lost the property. Plasmon resonance on matrix proteins and cellular impedance analyses showed in vitro that avidin exhibits a peculiar interaction with proteins and cells that allows the formation of highly stable Schiff's bases, after oxidation.
European Urology, 2003
Objective: To verify whether native avidin, made radioactive through the binding with technetium-99m labeled biotin (99m Tc-biotin), selectively accumulated in superficial tumor tissues following intravesical administration. Methodology: A total of fifteen patients with transitional cell bladder cancer were instilled intravesically with radiolabeled avidin. Cold biopsies were obtained from macroscopically normal and tumor tissues before transurethral resection (TUR) and the radioactivity in the samples was measured. Results: Increased accumulation of radiolabeled avidin was observed in tumor tissue compared to normal bladder tissue and in some cases, remarkably high quotients of uptake (q) in tumor versus normal tissues were determined (86 and 44). The three patients instilled with a deglycosylated avidin at neutral pI, who served as a control, showed no significant uptake in either tumor or normal urothelium and no difference in relative uptake (q ¼ 1:0). Conclusion: This pilot study indicated that intravesical administration of radiolabeled avidin resulted in a preferential accumulation in tumor tissue compared to normal urothelium. The instillation of radiolabeled avidin warrant further investigations in order to explore the possibility to treat superficial bladder neoplasms locally by replacing 99m Tc with high energy beta emitting radionuclides associated with biotin.
The Effect of Avidin on Viability and Proliferation of Colorectal Cancer Cells HT-29
Asian Pacific Journal of Cancer Prevention, 2022
Objective: The aim of this study was to analyze the effect of avidin treatment on cell viability, proliferation and cyclin D1 expression in colorectal cancer cells HT-29. Methods: Colorectal cancer cell line HT-29 incubated with 50, 100, 150, and 200 μg/mL of avidin concentration during 24, 48, and 72 hours, then the cell viability and proliferation were analyzed. Each avidin concentration was conducted together with HT-29 cell line without avidin treatment as a control group. The cell viability was measured by MTS assay and the proliferation was measured by BrdU (5-bromo-2′-deoxyuridine) cell proliferation assay. According to cell viability and proliferation result, we determined the 100 μg/ mL avidin concentration for analyzing mRNA and protein of cyclin D1. Results: We demonstrated that the viability and proliferation of HT-29 cells were significantly decreased in all concentration of avidin treatment compared to control. The cell proliferation showed larger reduction in avidin treatment rather than cell viability. This proves avidin could inhibit proliferation of colorectal cancer cell HT-29 quite well. The expression of cyclin D1, both mRNA and protein, was also significantly decreased after the avidin treatment group compared to control group, it supports the suppression of proliferation result. Conclusion: We concluded that avidin treatment could decrease cell viability and proliferation, accompanied by suppression of cyclin D1 expression in colorectal cells HT-29.
A spectrophotometric assay for biotinbinding sites of immobilized avidin
Applied Biochemistry and Biotechnology, 1996
A rapid and sensitive spectrophotometric assay was developed for the measurement of biotin-binding sites of immobilized avidin. The method is based on the reaction of avidin with excess biotin followed by assay of the unbound biotin using the HABA (2-[4'-hydroxyazobenzene]benzoic acid) method. Three solids possessing variable amounts of monomeric avidin were examined; viz., succinamidopropyl-controlled-pore glass (CPG-500), crosslinked 6% beaded agarose (Sepharose-CL-6B**), and crosslinked bis-acrylamide/azlactone (3M Emphaze Biosupport Medium AB1). Results indicate that the total biotin-binding sites of monomeric avidin immobilized on CPG-500, Sepharose-CL-6B, and 3M Emphaze are 0.229, 0.093, and 0.218/~m01 biotin per mL beads, respectively. Assays for exchangeable biotinbinding sites gave values greater than 90% of the total sites. The spectrophotometric HABA method described is an alternative to assays based on tracers, thus the handling of radioactive material is avoided.
Monoclonal Antibody and Avidin or Streptavidin
1996
Due to their high affinityfor biotin, avidin (Av) and streptavidin (SAy) are used to bridge pretargeted antibody molecules and radiolabeled biotin derivatives in vivo. Methods: We compared uptake of 125l@ labeled AyorSAv (— 1O-500 p@ g) intumorand normal tissues 3 days after a biotinylated B72. 3 monoclonal antibody (100@ &g) injection in nude mice. The animals were killed24 hrlater and the biodistribution of 125lwas determined. Results: The percent injected dose per gram of tumor remaIned constant over the range of injected ...
Targeted optical imaging of cancer cells using lectin-binding BODIPY conjugated avidin
Biochemical and Biophysical Research Communications, 2006
Lectins (asialo-receptor family) are expressed on a number of tumors that develop peritoneal metastases. To demonstrate that fluorescence imaging based on lectin binding is applicable for a variety of tumors, we conjugated BODIPY to avidin (avidin-BODIPY), and studied the efficacy of tumor targeting in 9 cancer cell lines in vitro and an ovarian cancer cell line in vivo using a murine peritoneal cancer model. All 9 cell lines showed specific intracellular accumulation with avidin-BODIPY on fluorescence microscopy and flow cytometry. In vivo spectral molecular imaging clearly visualized the peritoneal tumor foci with avidin-BODIPY, whereas, deglycosylated avidin-BODIPY (neutravidin-BODIPY) showed only minimal fluorescence from the tumor foci and was accompanied by higher background signals. These results suggest the lectin-targeted molecular imaging technique using a targeted green fluorescence probe is potentially useful in a wide variety of cancers with a proclivity for dissemination in the peritoneal space.
Amplified immunoperoxidase staining of isoelectrically focused human tumor markers
Electrophoresis, 1980
Amplified immunoperoxidase staining of isoelectrically focused human tumor markers Avidin-biotin amplified immunoperoxidase staining of the human tumor marker, carcinoembryonic antigen (CEA) localized in malignant tissue by isoelectric focusing is described. CEA either in electrofocused aqueous tissue extracts or extracted and separated by direct tissue isoelectric focusing in agarose (IsoGel), was reacted with rabbit anti-CEA antiserum and with enzyme conjugated swine anti-rabbit antiserum, or with avidin-biotin conjugated reagents, then with enzyme substrate. Avidin-biotin amplified immunoperoxidase staining of CEA gave more intense staining. Quantitation and characterization of tumor markers by direct tissue electrophoresis in isoelectric focusing media and enzyme amplification of antitumor marker antisera, make possible the study of their biological meaning and clinical usefulness.