scholarly journals What Is Limulus Amebocyte Lysate (LAL) and Its Applicability in Endotoxin Quantification of Pharma Products

2019 ◽  
Author(s):  
Yasir Mehmood
Keyword(s):  
Limulus Amebocyte Lysate

Interference of Gold Nanoparticles with In vitro Endotoxin Detection Assays

2020 ◽  
Vol 16 (2) ◽  
pp. 204-213 ◽  
Author(s):  
Melissa A. Vetten ◽  
Mary Gulumian
Keyword(s):  
Gold Nanoparticles ◽  
Gold Nanoparticle ◽  
Surface Reactivity ◽  
In Vitro Assays ◽  
Limulus Amebocyte Lysate ◽  
Chromogenic Assay ◽  
Nanoparticle Suspensions ◽  
Endotoxin Contamination ◽  
Factor C

Background: Endotoxin-free engineered nanoparticle suspensions are imperative for their successful applications in the field of nanomedicine as well as in the investigations in their toxicity. Gold nanoparticles are known to interfere with various in vitro assays due to their optical properties and potential for surface reactivity. In vitro endotoxin testing assays are known to be susceptible to interference caused by the sample being tested. Objective: This study aimed to identify a preferred assay for the testing of endotoxin contamination in gold nanoparticle suspensions. Methods: The interference by gold nanoparticles on three assays namely, the commonly used limulus amebocyte lysate chromogenic assay, the limulus amebocyte lysate gel-clot method, and the less common recombinant Factor C (rFC) assay, was tested. Results: Possible interference could be observed with all three assays. The interference with the absorbance- based chromogenic assay could not be overcome by dilution; whilst the qualitative nature of the gel-clot assay excluded the possibility of distinguishing between a false positive result due to enhancement of the sensitivity of the assay, and genuine endotoxin contamination. However, interference with the rFC assay was easily overcome through dilution. Conclusion: The rFC assay is recommended as an option for endotoxin contamination detection in gold nanoparticle suspensions.

scholarly journals Contamination of erythropoietin by endotoxin: in vivo and in vitro effects on murine erythropoiesis

Blood ◽  
1979 ◽  
Vol 54 (1) ◽  
pp. 146-158 ◽  
Author(s):  
KS Zuckerman ◽  
PJ Quesenberry ◽  
J Levin ◽  
R Sullivan
Keyword(s):  
Significant Inhibition ◽  
Erythropoietic Activity ◽  
Limulus Amebocyte Lysate ◽  
Endogenous Erythropoietin ◽  
Significant Change ◽  
In Vitro Effects ◽  
Stimulation Of

Abstract Endotoxin was detected in all erythropoietin preparations tested and was removed from four lots, without loss of erythropoietic activity, by adsorption with limulus amebocyte lysate. Comparison of adsorbed (endotoxin-depleted) and nonadsorbed (endotoxin-containing) erythropoietin preparations demonstrated significant inhibition of CFU- e and BFU-e in vitro by nonadsorbed erythropoietin at concentrations higher than 0.25 U/ml and 2.0 U/ml, respectively. CFU-e and BFU-e were inhibited significantly by readdition in vitro of 10(-5)-10(-3) mug of endotoxin per unit of limulus-adsorbed erythropoietin. Administration of saline or 6 U of nonadsorbed or adsorbed erythropoietin twice a day for 4 days of CF1 mice resulted in reticulocyte counts of 2.1%, 9.9%, and 15.9%, respectively. Nonadsorbed erythropoietin resulted in a 29% decrease in erythropoiesis, a 42% decrease in CFU-e, and a 16% increase in granulopoiesis in the marrow, whereas adsorbed erythropoietin caused a 28% increase in erythropoiesis, no significant change in CFU-e and a 19% decrease in granulopoiesis in the marrow. Both preparations resulted in marked increases in splenic erythropoiesis and granulopoiesis. The effects of adsorbed erythropoietin are similar to those produced following stimulation of hematopoiesis by endogenous erythropoietin. Hemopoietic changes induced by nonadsorbed erythropoietin in vivo and in vitro are affected substantially by contamination of the erythropoietin preparations with endotoxin.

scholarly journals Endotoxins in surgical instruments of hip arthroplasty

2016 ◽  
Vol 50 (3) ◽  
pp. 405-410 ◽  
Author(s):  
Vania Regina Goveia ◽  
Isabel Yovana Quispe Mendoza ◽  
Gilberto Lima Guimarães ◽  
Flavia Falci Ercole ◽  
Bráulio Roberto Gonçalves Marinho Couto ◽  
...  
Keyword(s):  
Femoral Head ◽  
Teaching Hospital ◽  
Hip Arthroplasty ◽  
Exploratory Study ◽  
Surgical Instruments ◽  
Gel Formation ◽  
Limulus Amebocyte Lysate ◽  
Positive Analysis ◽  
Hip Arthroplasties

Abstract OBJECTIVE To investigate endotoxins in sterilized surgical instruments used in hip arthroplasties. METHOD A descriptive exploratory study conducted in a public teaching hospital. Six types of surgical instruments were selected, namely: acetabulum rasp, femoral rasp, femoral head remover, chisel box, flexible bone reamer and femoral head test. The selection was based on the analysis of the difficulty in removing bone and blood residues during cleaning. The sample was made up of 60 surgical instruments, which were tested for endotoxins in three different stages. The EndosafeTM Gel-Clot LAL (Limulus Amebocyte Lysate method) was used. RESULT There was consistent gel formation with positive analysis in eight instruments, corresponding to 13.3%, being four femoral rasps and four bone reamers. CONCLUSION Endotoxins in quantity ≥0.125 UE/mL were detected in 13.3% of the instruments tested.

Endotoxin recovery using limulus amebocyte lysate (LAL) assay

Biologicals ◽  
2016 ◽  
Vol 44 (5) ◽  
pp. 434-440 ◽  
Author(s):  
Jay S. Bolden ◽  
Rob E. Warburton ◽  
Robert Phelan ◽  
Marie Murphy ◽  
Kelly R. Smith ◽  
...  
Keyword(s):  
Limulus Amebocyte Lysate ◽  
Lal Assay

Application of quartz tuning forks for detection of endotoxins and Gram-negative bacterial cells by monitoring of Limulus Amebocyte Lysate coagulation

2014 ◽  
Vol 58 ◽  
pp. 132-137 ◽  
Author(s):  
Andrzej Chałupniak ◽  
Karol Waszczuk ◽  
Katarzyna Hałubek-Głuchowska ◽  
Tomasz Piasecki ◽  
Teodor Gotszalk ◽  
...  
Keyword(s):  
Bacterial Cells ◽  
Limulus Amebocyte Lysate ◽  
Gram Negative ◽  
Tuning Forks ◽  
Quartz Tuning Forks

Evaluation of absorption with Limulus amebocyte lysate to remove contaminating endotoxin from interferon and lymphokine preparations

1984 ◽  
Vol 66 (1) ◽  
pp. 103-112 ◽  
Author(s):  
A. Biondi ◽  
S. Landolfo ◽  
D. Fumaroa ◽  
N. Polentarutti ◽  
M. Introna ◽  
...  
Keyword(s):  
Limulus Amebocyte Lysate

Endotoxins in Ophthalmic Viscosurgical Devices

2003 ◽  
Vol 13 (2) ◽  
pp. 176-184 ◽  
Author(s):  
H. Burkhard Dick ◽  
A.J. Augustin ◽  
T. Pakula ◽  
N. Pfeiffer
Keyword(s):  
Positive Sample ◽  
Gram Negative Bacteria ◽  
European Pharmacopoeia ◽  
Limulus Amebocyte Lysate ◽  
Dilution Series ◽  
Sample Extract ◽  
Immunological Reactions ◽  
Negative Controls ◽  
Lal Assay

Purpose To measure the endotoxin concentration (EC) of 25 commercially available, hyaluronic acid- and hydroxypropylmethylcellulose-based (HPMC) ophthalmic viscosurgical devices (OVDs). Methods The in vitro Limulus amebocyte lysate (LAL) assay, which indicates the presence of endotoxins originating from gram-negative bacteria, was used to determine the EC. The procedure was performed according to the European Pharmacopoeia/USP. EC including duplicate determinations, negative controls, dilution series with control standard endotoxin, dilution series with sample extract and positive sample control. Results 16 OVDs (Amvisc®, Amvisc® Plus, Biolon®, Coatel®, Healon®, Healon® GV, Healon®5, HPMC Ophtal® L, Microvisc®, Microvisc® Plus, Ocucoat®, Provisc®, Rayvisc®, Viscoat®, Visco Shield® 2%, Visko® 1.4%) had an EC under 1.2 endotoxin units/mL, five (Adatocel®, HPMC Ophtal® H, LA Gel®, Viscorneal®, Viscorneal® Plus) had an EC ≥ 1.2 and ≤ 24 EU/ml, and four (Biocorneal®, Dispasan® also named Ophthalin, Dispasan® Plus, Visko® 1%) had an EC of > 24 EU/ml. Discussion To avoid viscoelastic-related inflammatory or immunological reactions, the use of pure OVDs is recommended, especially for surgical procedures with an inherent possibility of leaving viscoelastic remnants in the eye (e.g., cataract surgery, viscocanalostomy or penetrating keratoplasty).

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