Sponge‐Like: A New Protocol for Preparing Bacterial Ghosts

Abstract

Bacterial Ghosts (BGs) received an increasing interest in the recent years for their promising medicinal and pharmaceutical applications. In this study, for the first time we introduce a new protocol for BGs production. E. coli BL21 (DE3) pLysS (Promega) was used as a model to establish a general protocol for BGs preparation. The protocol is based on using active chemical compounds in concentrations less than the Minimum Inhibition Concentration (MIC). Those chemical compounds are SDS, NaOH, and H2O2. Plackett-Burman experimental design was used to map the best conditions for BGs production. Normal and electronic microscopes were used to evaluate the BGs quality (BGQ). Spectrophotometer was used to evaluate the amount of the released protein and DNA. Agarose gel electrophoresis was used to determine the existence of any residue of DNA after each BGs preparation. Viable cells, which existed after running this protocol, were subjected to lysis by inducing the lysozyme gene carried on pLysS plasmid. This protocol is able to produce BGs that can be used in different biotechnological applications.

Vaccines against Pathogens: A Review and Food For Thought

“…Apparently, the first attempt to design a full protocol to introduce pore(s) in bacterial cells using the concept of using critical chemical concentration (both of the MIC and the MGC) of some compounds which did not damage the cells 3D structure under the used concentration were Amara, et al [66].…”

Lysozymes, Proteinase K, Bacteriophage E Lysis Proteins, and some Chemical Compounds for Microbial Ghosts Preparation: a Review and Food for Thought

“…After enough washing and centrifugation steps, 60 percent ethanol is used in the third step. Each of the twelve used experiments is following the Plackett-Burmen design as in the original protocol [41].…”

Cell Evacology: Could it be a New Science

“…The sponge like protocol successfully introduced a fast, cheap and in house cell evacuation for nearly all the biological cells [1]. The original protocol describes bacterial cell evacuation from its cytoplasmic contents [1,2].…”

“…The original protocol describes bacterial cell evacuation from its cytoplasmic contents [1,2]. The protocol was optimized using Plackett-Burman design, which enables randomization for the variables included.…”

“…The protocol was optimized using Plackett-Burman design, which enables randomization for the variables included. Such randomization enables mapping the best cells' evacuation experimental conditions, which give the best un-deformed 3D structure and un-deteriorated surface antigens [1,3]. Its main concept is to determine the minimum inhibition concentration (MIC) and the minimum growth concentration (MGC) of chemical compounds able to induce pores in the cell walls [1,3].…”