arginyl-glycyl-aspartyl-phenylalanine and 1-4-7-10-tetraazacyclododecane--1-4-7-10-tetraacetic-acid

A robust, versatile and compact microreactor has been designed, fabricated and tested for the labeling of bifunctional chelate conjugated biomolecules (BFC-BM) with PET radiometals.. The developed microreactor was used to radiolabel a chelate, either 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) or 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA) that had been conjugated to cyclo(Arg-Gly-Asp-DPhe-Lys) peptide, with both ⁶⁴Cu and ⁶⁸Ga respectively. The microreactor radiolabeling conditions were optimized by varying temperature, concentration and residence time.. Direct comparisons between the microreactor approach and conventional methods showed improved labeling yields and increased reproducibility with the microreactor under identical labeling conditions, due to enhanced mass and heat transfer at the microscale. More importantly, over 90% radiolabeling yields (incorporation of radiometal) were achieved with a 1:1 stoichiometry of bifunctional chelate biomolecule conjugate (BFC-BM) to radiometal in the microreactor, which potentially obviates extensive chromatographic purification that is typically required to remove the large excess of unlabeled biomolecule in radioligands prepared using conventional methods. Moreover, higher yields for radiolabeling of DOTA-functionalized BSA protein (Bovine Serum Albumin) were observed with ⁶⁴Cu/⁶⁸Ga using the microreactor, which demonstrates the ability to label both small and large molecules.. A robust, reliable, compact microreactor capable of chelating radiometals with common chelates has been developed and validated. Based on our radiolabeling results, the reported microfluidic approach overall outperforms conventional radiosynthetic methods, and is a promising technology for the radiometal labeling of commonly utilized BFC-BM in aqueous solutions.

Topics: Copper Radioisotopes; Dimethylpolysiloxanes; Gallium Radioisotopes; Glass; Heterocyclic Compounds; Heterocyclic Compounds, 1-Ring; Isotope Labeling; Microfluidic Analytical Techniques; Oligopeptides; Positron-Emission Tomography; Temperature; Time Factors

We report the design and synthesis of a trifunctional probe for seeing and counting cancer cells using both fluorescence imaging (FI) and inductively coupled plasma mass spectrometry (ICPMS) for the first time. It consisted of a guiding cyclic RGD peptide unit to catch cancer cells via targeting the α(v)β(3) integrin overexpressed on their surface, a 5-amino-fluorescein moiety for FI using confocal laser scanning microscopy (CLSM) as well as a 2-aminoethyl-monoamide-DOTA group for loading stable europium ion and subsequent ICPMS quantification of the cancer cells without the use of radioactive isotopes. In addition to FI, the LOD (3σ) of the α(v)β(3) integrin was down to 69.2-309.4 amol per cell depending on the type of the α(v)β(3)-positive cancer cells when using ICPMS and those of the cancer cell number reached 17-75. This probe developed enables us not only to see but also to count the α(v)β(3)-positive cancer cells ultrasensitively, paving a new way for early diagnosis of cancer.

Topics: Amides; Cell Count; Cell Line, Tumor; Drug Design; Europium; Fluorescein; Fluorescent Dyes; Heterocyclic Compounds, 1-Ring; Humans; Integrin alphaVbeta3; Molecular Probes; Neoplasms; Oligopeptides