Biomolecular imaging of colorectal tumor lesions using a FITC-labeled scFv-Cκ fragment antibody

Abstract

For the sensitive diagnosis of colorectal cancer lesions, advanced molecular imaging techniques using cancer-specific targets have emerged. However, issues regarding the clearance of unbound probes and immunogenicity remain unresolved. To overcome these limitations, we developed a small-sized scFv antibody fragment conjugated with FITC for the real-time detection of colorectal cancer by in vivo molecular endoscopy imaging. A small-sized scFv fragment can target colon cancer secreted protein-2 (CCSP-2), highly expressed in colorectal adenocarcinoma tissues; moreover, its full-length IgG probe has been used for molecular imaging previously. To assess the efficacy of anti-CCSP-2 scFv-FITC, surgical specimens were obtained from 21 patients with colorectal cancer for ex vivo molecular fluorescence analysis, histology, and immunohistochemistry. Orthotopic mice were administered with anti-CCSP-2 scFv-FITC topically and intravenously, and distinct tumor lesions were observed by real-time fluorescence colonoscopy. The fluorescence imaging of human colon cancer specimens allowed the differentiation of malignant tissues from non-malignant tissues (p < 0.05), and the CCSP-2 expression level was found to be correlated with the fluorescence intensity. Here, we demonstrated the feasibility and safety of anti-CCSP-2 scFv-FITC for molecular imaging as well as its potential in real-time fluorescence colonoscopy for the differential diagnosis of tumor lesions.

Feasibility of moxifloxacin and proflavine dual fluorescence imaging for detecting gastrointestinal neoplastic lesions: A prospective study

“…In this regard, molecular imaging using specific targeting probes with wide-field endoscope would be promising with molecular and functional information, although it has not been widely used until now. [22][23][24][25][26] In the era of personalized and precision medicine, endoscopy providing real-time information about the exact tumor size, histopathological characteristics associated with poor prognosis such as lymphovascular or perineural invasion which is associated with increased possibility of lymph node metastasis, is also promising. Thus, more specialized high-contrast and high-resolution imaging methods would be helpful for the sensitive detection of gastrointestinal lesions.…”

“…Regarding the image contrast of CLE, fluorescein cannot stain cells thus cells appear in negative contrasts. In this regard, molecular imaging using specific targeting probes with wide‐field endoscope would be promising with molecular and functional information, although it has not been widely used until now 22–26 . In the era of personalized and precision medicine, endoscopy providing real‐time information about the exact tumor size, histopathological characteristics associated with poor prognosis such as lymphovascular or perineural invasion which is associated with increased possibility of lymph node metastasis, is also promising.…”

Investigation of artificial intelligence integrated fluorescence endoscopy image analysis with indocyanine green for interpretation of precancerous lesions in colon cancer

“…Fluorescence-based tumor imaging can be a useful technique for early cancer detection owing to its advantageous high specificity and sensitivity, real-time imaging, and absence of radiation exposure [ 8 ]. In previous studies, a fluorescence-labeled compound, which could bind to tumor-specific molecules, such as vascular endothelial growth factor (VEGF) [ 9 , 10 ], epidermal growth factor receptor (EGFR) [ 11 ], colon cancer specific protein-2 (CCSP-2) [ 12 ], endothelial A receptor (ET A R) [ 13 ], and c-Met [ 14 ] was developed to detect colon cancer. Although various probes have been used, their safety in the human body and their ability to detect precancerous lesions, such as adenoma, serrated polyps, and dysplasia, must be confirmed before applying this technology in clinical endoscopic examination [ 15 ].…”

Machine learning modeling for solubility prediction of recombinant antibody fragment in four different E. coli strains

“…Its molecular weight is considerably smaller than the full-length antibodies. Owing to small size and low immunogenicity, scFv has brought much attention in biomedicine for theranostic purposes 11 . 4D5MOC-B scFv is a stable anti EpCAM extracellular domain-scFv (anti EpEX-scFv) with a very high affinity to its target.…”