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| Author and year | Study title | Study findings | Reference(s) |
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| Bolden and Smith [106] | Application of recombinant factor C reagent for the detection of bacterial endotoxins in pharmaceutical products | In this study, the rFC reagent was used as part of an endpoint florescence-based endotoxin method to test a number of pharmaceutical products. The method is the same as or better than the bacterial endotoxin test (BET) method in the compendia. | [106] |
| Schwarz et al. [107] | Residual endotoxin contaminations in recombinant proteins are sufficient to activate human CD1c+ dendritic cells | Endotoxin units (EU) are commonly used to measure the amount of bacterial contamination in commercially produced recombinant proteins. Most suppliers guarantee less than 1 EU level, roughly equivalent to 100 pg of E. coli LPS per microgram of recombinant protein. 1-100 pg LPS may be present in protein preparations with 10-1000 ng/ml values. The current study examines the effects of extremely low endotoxin concentrations ranging from 0.002 to 2 ng/ml on human immune cells because the majority of in vitro research has concentrated on endotoxin effects produced by concentrations between 1 and 100 ng/ml. | [107] |
| Reich et al. [108] | Detection of naturally occurring bacterial endotoxins in water samples | In this study, three commercially available synthetic reagents showed a correlation of 94.4% when evaluating water from different sources. | [108] |
| Chen and Mozier [109] | Comparison of Limulus amebocyte lysate test method for endotoxin measurement in protein solutions | The kinetic turbidimetric approach, the kinetic chromogenic method, the endpoint chromogenic method, and the new recombinant factor C method (PyroGene) were all compared to the LAL test. The endotoxin test allows for a 2-fold error (50-200%) and a spike recovery of 50-200%. To rule out interfering molecules, such as LPS interaction with proteinaceous or lipophilic chemicals, this spike recovery research was done on all samples in the test plates. This practice is beneficial, but exact precision at low levels is difficult to determine. The majority of endotoxin values fall within a 2-fold range. Some samples, such as yeast samples, may require pretreatment before testing. | [109] |
| Grallert et al. [110] | EndoLISA®: a novel and reliable method for endotoxin detection | In this study, the primary receptor of the Limulus amebocyte coagulation cascade reacts with factor C to detect LPS. The detection range of EndoLISA (rFC) is 0.05 EU/ml to 500 EU/ml. | [110] |
| McKenzie et al. [111] | Evaluation of lot-to-lot repeatability and effect of assay media choice in the recombinant factor C assay | This study demonstrated that rFC assays of environmental materials are repeatable from lot to lot and that employing Tris buffer or water as the extraction and assay medium for endotoxin assessment in dust samples may be a viable choice for the development of a standardized method. | [111] |
| Thorne et al. [112] | Evaluation of the Limulus amebocyte lysate and recombinant factor C assays for assessment of airborne endotoxin | The results of this study show that the LAL and rFC assays deliver the same amount of information about how much endotoxin is in the air in livestock facilities. | [112] |
| Loverock et al. [113] | A recombinant factor C procedure for the detection of Gram-negative bacterial endotoxin | A photometric approach for detecting Gram-negative bacterial endotoxins is described in this article (lipopolysaccharide). The traditional Limulus amebocyte lysate (LAL) cascade is initiated by the activation of recombinant factor C (rFC). The rFC technique described here measures the cleavage of a fluorogenic substrate by an enzyme. | [113] |
| Bolden et al. [114] | Results of a harmonized endotoxin recovery study protocol evaluation by 14 BioPhorum Operations Group (BPOG) member companies | A quantitative fluorometer is used in the endpoint fluorescence assay. Recombinant factor C, an endotoxin-sensitive protein, a fluorogenic substrate, and a buffer are used in this experiment. Fluorescence is analyzed at the beginning of the investigation and after 60 or 90 minutes of incubation. The variation in fluorescence is therefore proportional to the endotoxin concentration. The samples were diluted and then examined in accordance with site protocols. Diluted samples were mixed with recombinant factor C, a fluorogenic substrate, and buffer in a 96-well microplate. The endotoxin concentration was ascertained by contrasting the fluorescence variation after an hour of incubation with the standard curve. Linear regression was used to calculate the results. | [114] |
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