Labeling of Anti-MUC-1 Binding Single Chain Fv Fragments to Surface Modified Upconversion Nanoparticles for an Initial in Vivo Molecular Imaging Proof of Principle Approach

Abstract

In vivo optical Imaging is an inexpensive and highly sensitive modality to investigate and follow up diseases like breast cancer. However, fluorescence labels and specific tracers are still works in progress to bring this promising modality into the clinical day-to-day use. In this study an anti-MUC-1 binding single-chain antibody fragment was screened, produced and afterwards labeled with newly designed and surface modified NaYF 4 :Yb,Er upconversion nanoparticles as fluorescence reporter constructs. The MUC-1 binding of the conjugate was examined in vitro and in vivo using modified state-of-the-art small animal Imaging equipment. Binding of the newly generated upconversion nanoparticle based probe to MUC-1 positive cells was clearly shown via laser scanning microscopy and in an initial proof of principal small animal optical imaging approach.

Selecting Targets for Tumor Imaging: An Overview of Cancer-Associated Membrane Proteins

“…Although relatively few considering the NCI rating, several approaches are being evaluated to target MUC-1 therapeutically, including antibodies (fragments), peptides, and nucleic acid aptamers, 97 , 98 some of which are also under evaluation for various imaging applications. 99 – 101 …”

“…Although relatively few considering the NCI rating, several approaches are being evaluated to target MUC-1 therapeutically, including antibodies (fragments), peptides, and nucleic acid aptamers, 97,98 some of which are also under evaluation for various imaging applications. 99–101 Extracellular matrix metalloproteinase inducer (EMMPRIN), also named basigin or CD147, is another example of a type I transmembrane glycoprotein that does not fit within any category based on its function. EMMPRIN acts as a stimulator of MMP synthesis via cell-cell interactions.…”

Polymer-antibody fragment conjugates for biomedical applications

“…This chemoselective reaction can occur under physiological conditions and involves initial formation of a thioester-linked species by reaction between N-and C-terminal cysteine residues on two different protein molecules, followed by spontaneous rearrangement of the thioester-linked species via an intramolecular reaction to form a native peptide bond at the ligation site (21). A variety of studies have used NCL for the bioconjugation of oligopeptides and recombinant proteins to dendrimers [173,174] as the approach allows chemoselective dendrimer conjugation to the terminus of a protein without interfering with protein function.…”

“…Isolated antibodies can be readilyaffinity maturedto improve their binding and engineered to create smaller, antibody-derived fragments of <10 nm dimensions that can be further modified to facilitate extensive chemical functionalization. This has led to increasing encroachment of fragments into lucrative fields traditionally associated with monoclonal antibody-polymer conjugates, such as therapeutics where humanized and PEGylatedantibody fragments have gained regulatory approval in recent yearsfor treatment of Crohn's disease and rheumatoid arthritis ( [15]; reviewed in [16]), and immunodetection, where small, easily functionalized fragments are fast becoming the molecules of choice for in vitro detection and monitoring applications [17][18][19][20] and, somewhat more cautiously, in vivoimaging and diagnostics [21,22].…”

Noncontact recognition of fluorescently labeled objects in deep tissue via a novel optical light beam arrangement

“…In currently available optical imaging systems, high fluorescent dye concentrations and/or a high laser power combined with a sensitive low-noise charge-coupled device (CCD) or complementary metal-oxide-semiconductor (CMOS) detectors are required for the imaging of deep tissue layers [10,25,28]. However, the required high excitation intensity can cause tissue overheating, leading to serious damage [29]. Thus, light in the infrared range is preferable because it is less scattered and absorbed by tissue than light in the visible spectrum, while simultaneously providing sufficient emission signals.…”