New electroporation protocols for Cargo Loading
The research group of Biopulse Biotech is working on the development of innovative physico-chemical methods that allow the loading of molecules of various kinds in nanovesicles. We specialize in the development of electrical impulses and applications with electroporation methods. For this reason, our team is applying know-how to find new electroporation protocols dedicated to Cargo Loading technologies. Our research in cargo loading technologies is focused on finding efficient ways to help scientists easily load liposomes, exosomes or new types of plant nanovesicles such as plant exosomes or algosomes. The goal of our electroporation tests is to set reliable protocols for new Cargo Loading technologies in terms of retention capacity and stability of the nanovesicle.
The goal of our research group is to transmit sufficient scientific information to the R&D department to develop an innovative electroporation technology. Based on the tests conducted, in fact, we are able to provide researchers with medical devices that perform functions of loading nanovesicles and experiments on laboratory animals in a completely flexible way and in a single product.
As for example with our BioDischarger electroporator®, able to electroporate in vitro and in vivo with extreme flexibility. In fact, the fields of application of our electroporators for Cargo Loading technology, can range from in vitro loading of cell cultures, exosomes and liposomes, to in vivo work of experimental tumors on animal models and on mouse muscle tissue by injecting drugs, oligonucleotides and plasmid DNA. In the case of the BioDischarger® electroporator it is possible to carry out in vitro and in vivo operations with a single instrument that is able to set the characteristics of the different electrical impulses according to the model and loading characteristics. As part of our research on electroporation applied to Cargo Loading technologies, we have carried out some tests on innovative electrical pulse protocols to charge nanovesicles extracted from plants.
The BioDischarger®
The Biopulse Biotech research group has in fact focused on development tests of technologies that allow to satisfactorily load, through the BioDischarger® electroporator inside the nanovesicles, for example vitamins, substances for cosmetic use, drugs and agents for medical, veterinary pharmacological and food use. Since the first use of the hardware prototype in the tests, the electroporator developed by Biopulse Biotech, intended for cargo loading technology, has proven to be efficient and extremely versatile. In fact, our electroporator has allowed the loading of plant nanovesicles with a physico-chemical method, through the transient destabilization of these nanovesicles. During the tests we used different combinations of single electrical pulses and as a wavetrain, with variable voltage and duration, so that we always use the most efficient sequence for loading.
During the tests, the applied wavetrains induced a transient permeabilization of the plant nanovesicle or exosome, which then returned, after a variable time and in any case less than thirty minutes, to the initial stage of electrical inertia. This period of permealization effectively led to increased transmembrane traffic that allowed exosomal loading with various molecules depending on the purpose needed. On our BioDischarger electroporators®, electrical and chemical-physical parameters are constantly measured during permeabilization and loading in order to improve the efficiency of the procedure.
Our Cargo Loading tests
The R&D department of Biopulse Biotech, has built several hardware prototypes, to test different configurations of plant exosomes uploads during the Cargo Loading tests. For example, different universal bases were created to accommodate cuvettes of various kinds, and other complementary accessories that helped our scientists to measure certain parameters during loading tests, such as temperature and electrical impedance during applied pulses. In the laboratories we have loaded different substances inside the plant exosomes, with excellent results in terms of retention, especially when compared to the tests carried out on liposomes. The molecules of various nature were loaded into isolated suspended and resuspended plant nanovesicles analyzed with Nanoparticle Tracking Analysis using Nanosight for the evaluation of concentration and size distribution.
All the prototypes and evolutions of the BioDischarger electroporators® built by Biopulse Biotech, have led our research group to conclude the first settings of the electroporators intended for the market. Normally, the basic electrical protocol provides for the first electrical impulse with the function of destabilizing the membrane making it permeable, and the subsequent train of electrical pulses to facilitate the transmembrane ionic (or molecular) transit. All the presets of electrical protocols applicable to Cargo Loading technology are completely expandable and customizable in terms of duration and electrical voltage. Our research group has allowed us to install a broth of the BioDischarger electroporator®, a series of durations and voltages that take into account the different sizes and stability of exosomal membranes, which have been developed on the basis of experiences of conveying molecules inside cells.
