scholarly journals Pyrogen testing revisited on occasion of the 25th anniversary of the whole blood monocyte activation test

ALTEX ◽  
2021 ◽  
pp. 3-19
Author(s):  
Thomas Hartung
Keyword(s):  
Whole Blood ◽  
Implementation Process ◽  
Monocyte Activation ◽  
Limulus Amebocyte Lysate ◽  
25Th Anniversary ◽  
Safety Testing ◽  
Cell Therapies ◽  
Horseshoe Crabs ◽  
One Year ◽  
Lal Assay

The whole blood pyrogen test invented 25 years ago, and its variant based on cryo-preserved blood one year later, brought momentum into the field of pyrogen testing, which, despite the broad application of the Limulus amebocyte lysate (LAL) assay, aka bacterial endotoxin test (BET), consumed several hundred thousand rabbits per year world-wide. The resulting international validation and lengthy acceptance and implementation process of what are called now monocyte activation tests (MATs) finally is impacting on animal numbers – at least in Europe – reducing them by more than 70% and counting. The author sees no reason for continuing any regulatory rabbit testing for pyrogens except the lack of acceptance of MATs in some regions of the world. The availability of MATs has opened also the discussion about the shortcomings of LAL/BET, namely its restriction to Gram-negative pyrogens, non-reflection of the potency of these in humans, interference and masking by many products, and animal welfare concerns for horseshoe crabs. The obvious advantages of MATs in all these respects should lead to a shift from LAL/BET to MATs. We are starting to see this for vac­cines and medical devices, but other areas like safety testing of blood transfusions, cell therapies and nanomaterials, and the assessment of air-borne pyrogens still need to grasp the opportunity provided by MATs. While the different MATs can jointly serve these needs, the whole blood MAT has some advantages as discussed here.

Regulating Innovation in the Early Development of Cell Therapies

2020 ◽  
Author(s):  
Andrew R Exley ◽  
James McBlane
Keyword(s):  
Clinical Trials ◽  
Nk Cell ◽  
Genetic Manipulation ◽  
Clinical Trial Data ◽  
Cell Receptor ◽  
Mitigation Measures ◽  
Safety Testing ◽  
Cell Therapies ◽  
Commercial Use ◽  
Allogeneic Cell

Abstract Clinical need for paradigm shifts in efficacy and safety is driving the rapid and wide-ranging innovation in cell therapies for cancer beyond existing regulatory frameworks. Critical issues emerging during clinical trials frequently reflect unresolved elements of the regulation of innovation conundrum from earlier stages of development. We address this challenge using a global regulators’ perspective on the pre-clinical development of cell therapies, as a navigational aid to intended commercial use which maximises the clinical relevance of developmental data. We examine the implications of tumour targeting based on B cell, NK cell, conventional and unconventional T cell receptor domains; multiplex approaches; genetic manipulation strategies; and autologous versus allogeneic cell sources. We propose that detailed characterisation of both the cell source and final product is critical to optimising manufacture of individualised autologous or off the shelf allogeneic cell therapies, enabling product consistency to underpin extrapolation of clinical trial data to the expected commercial use. We highlight preclinical approaches to characterising target antigens including the Human Cell Atlas initiative, multi-dimensional cell culture, and safety testing against activated, proliferating or stressed control cells. Practical solutions are provided for preclinical toxicity studies when cell therapies target uniquely human tumour antigens, including illustrative mitigation measures for potential toxicity likely to support timely approval of first in human clinical trials. We recommend addressing the regulation of innovation conundrum through serial engagement between innovators and regulators early in the development of cell therapies for cancer, accelerating patient access whilst safeguarding against unacceptable toxicities.

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

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).

scholarly journals Evaluation of a Whole-Blood Cytokine Release Assay for Use in Measuring Endotoxin Activity of Group B Neisseria meningitidis Vaccines Made from Lipid A Acylation Mutants

2009 ◽  
Vol 17 (1) ◽  
pp. 98-107 ◽  
Author(s):  
Mark B. Stoddard ◽  
Valerian Pinto ◽  
Paul B. Keiser ◽  
Wendell Zollinger
Keyword(s):  
Neisseria Meningitidis ◽  
Whole Blood ◽  
Lipid A ◽  
Mononuclear Cells ◽  
Membrane Vesicle ◽  
Cytokine Release ◽  
Tnf Α ◽  
Release Assay ◽  
Group B ◽  
Lal Assay

ABSTRACT Bacterial endotoxin interacts with the human immune system via complex immunological pathways. The evaluation of endotoxicity is important in the development of safe vaccines and immunomodulatory therapeutics. The Limulus amebocyte lysate (LAL) assay is generally accepted by the FDA for use for the quantification of lipopolysaccharide (LPS), while the rabbit pyrogen test (RPT) is used to estimate pyrogenicity during early development and production. Other in vitro assays, such as cytokine release assays with human whole blood (WB) or peripheral blood mononuclear cells (PBMCs), have also been used and may better estimate the human immunological response to products containing novel LPS molecules. In this study, WB and PBMC interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) release assays were used to estimate the endotoxic activities of purified LPS and native outer membrane vesicle (NOMV) vaccines derived from wild-type (hexa-acylated lipid A) and genetically detoxified (penta- and tetra-acylated lipid A) group B Neisseria meningitidis. A method for quantification of the differences in endotoxicity observed in the WB and PBMC assays is elucidated. The LAL assay was shown to be relatively insensitive to lipid A variations, and the RPT was less sensitive than the cytokine release assay with WB. The IL-6 and TNF-α assays with WB but not the assays with PBMCs distinguished between vaccines containing LPS from penta- and tetra-acylated strains. The high degree of sensitivity of the WB system to LPS variations and the presumed relevance of the use of human tissues to predict toxicity in humans suggest that this assay may be particularly well suited for the safety evaluation of vaccines and therapeutics containing acylation variants of LPS.

scholarly journals Is Platelet-rich plasma superior to whole blood in the management of chronic tennis elbow: one year randomized clinical trial

2014 ◽  
Vol 6 (1) ◽  
Author(s):  
Seyed Ahmad Raeissadat ◽  
Seyed Mansoor Rayegani ◽  
Hossein Hassanabadi ◽  
Rosa Rahimi ◽  
Leyla Sedighipour ◽  
...  
Keyword(s):  
Clinical Trial ◽  
Randomized Clinical Trial ◽  
Whole Blood ◽  
Tennis Elbow ◽  
Platelet Rich Plasma ◽  
One Year

scholarly journals The un-infectivity of the PF cultivated strain of trypanosoma cruzi to mice. An evaluation through a one year period by blood cultures and histopathology

1975 ◽  
Vol 9 (1) ◽  
pp. 1-5 ◽  
Author(s):  
Humberto Menezes
Keyword(s):  
Trypanosoma Cruzi ◽  
Whole Blood ◽  
Blood Cultures ◽  
Subcutaneous Route ◽  
One Year

Trypanosoma cruzi of the cultivated PF strain when injected in mice per subcutaneous route, in adequate doses, is able to induce an efficient sterile immunization in the animais (for at least one year) as determined by whole blood cultures and histopathology.

Sign in / Sign up

Export Citation Format

Share Document