To date, generation of single-chain fragment variable (scFv) has become an established technique used to produce a completely functional antigen-binding fragment in bacterial systems. The advances in antibody engineering have now facilitated a more efficient and generally applicable method to produce Fv fragments. Basically, scFv antibodies produced from phage display can be genetically fused to the marker proteins, such as fluorescent proteins or alkaline phosphatase. These bifunctional proteins having both antigen-binding capacity and marker activity can be obtained from transformed bacteria and used for one-step immunodetection of biological agents. Alternatively, antibody fragments could also be applied in the construction of immunotoxins, therapeutic gene delivery, and anticancer intrabodies for therapeutic purposes. This paper provides an overview of the current studies on the principle, generation, and application of scFv. The potential of scFv in breast cancer research is also discussed in this paper.
Spheroids are generally self-assembled cells with the ability to generate their extracellular matrix, including the complex cell-matrix and the cell-cell interactions that resemble the functional characteristics of the corresponding tissue in vivo. The study aimed to develop a three-dimensional (3D) spheroid system for the cervical cancer cell lines, HeLa (HPV18), CaSki (HPV16), SiHa (HPV16), C33A (non-HPV), HT3 (non-HPV) as well as to identify its biological activity in the extracellular form. For the formation of the cervical cancer spheroids, the liquid overlay approach was applied, followed by embedding to the bovine collagen I matrix. Spheroid formation using the liquid overlay approach is achieved by growing the cells on a non-adhesive surface to prevent cellular adhesion, resulting in the cells forming aggregates and, subsequently, the spheroids. The obtained data shows the definite biological behavior of each of the particular cervical cancer cell lines, indicating that cells adapt their natural phenotype in a three-dimensional microenvironment.
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