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Human adenovirus type 5 Reference Material

Culture comparison of Human adenovirus type 5 (ATCC VR-5) and Human adenovirus type 5 reference material (ATCC VR-1516)

All stocks of the Human adenovirus type 5 Reference Material (ATCC VR-1516) have been depleted. The Adenovirus Reference Material Working Group (ARMWG) is currently working with ATCC, the National Institute of Standards and Technology, and industry partners to produce more of this material; the ISBioTech website provides updates as available. 

We have found that researchers have begun purchasing Human adenovirus type 5 (ATCC VR-5) in lieu of the Human adenovirus type 5 Reference Material (ATCC VR-1516) during this time. While the Human adenovirus type 5 Reference Material (ATCC VR-1516) was developed from Human adenovirus type 5 (ATCC VR-5), it must be noted that Human adenovirus type 5 (ATCC VR-5) is not highly purified or as extensively characterized as the reference material and thus may not be suitable for all applications. We have provided a comparison below for reference. 

 

Table 1. Culture comparison of Human adenovirus type 5 and Human adenovirus type 5 reference material

  Human adenovirus type 5 Human adenovirus type 5 Reference material
Catalog Number ATCC VR-5   ATCC VR-1516
Description Fluid and cell lysate from infected cultures Purified Adenovirus Type 5 (ATCC VR-5) formulated as sterile liquid in 20 mM TRIS, 25 mM NaCl, 2.5% glycerol, pH 8.0
Origin Spontaneously degenerating tissue culture of adenoid tissue from a 4-year-old girl with enlarged chronically infected tonsils, Washington, DC, 1953 Virus Bank derived from ATCC VR-5 (wild type)
Host cells HeLa (ATCC CCL-2 HEK 293 (ATCC CRL-1573)
Infectious titer Specification: ≥ 5×103 TCID50/mL
The most recent lot was 1.1×109 TCID50/mL in 7 days on HeLa cells, as determined by immunofluorescence.
7×1010 NIU/mL in HEK 293 cells, with 95% certainty that the range lies within 7×1010 to 8×1010 NIU/mL, based on data submitted from 17 different laboratories.
Particle concentration  Not tested  5.8×1011 particles/mL, with 95% certainty that the range lies within 5.6×1011 to 6.0×1011 particles/mL, as determined by A 260nm/SDS procedure, based on data submitted from 13 different laboratories.
Electron microscopy  Not tested Confirmed >70% of the material consists of icosahedral shaped single adenovirus particles, with the remaining population being doublets, triplets or multiplets consisting of more than three adenovirus particles.
Authentication
Genomic authentication was performed by NGS and results meet requirements of ≥ 1,000 bp of target gene with ≥ 98% homology to GenBank accession number AC_000008.1 (Human adenovirus 5, complete genome). The complete DNA sequence was determined and is deposited with GenBank. The ARM GenBank accession number is AY339865. The sequence is similar, but not identical, to the adenovirus 5 sequence found in GenBank accession number NC_001406 (Human adenovirus 5, complete genome).
Sterility Test for Mycoplasma Contamination: Broth and agar culture (direct method) DNA detection by PCR of test article nucleic acid
General Sterility test (BacT/ALERT 3D):
iAST bottle (aerobic) at 32.5°C, 14-day incubation
iNST bottle (anaerobic at 32.5°C, 14-day incubation
Tested and found free of adventitious agents.
Endotoxin was < 0.15 EU/mL.
Residuals  Not tested Less than 3 pg of residual host cell DNA per µg of total DNA. Residual HEK 293 host cell proteins were found to be 18 ng/mL. Residual BSA was found to be <0.5 ng/mL. there is approximately 2.0 µg of free hexon per 1012 adenoviral particles. No 31K MW precursor protein form was found. The A260nm to A280nm ratio in the presence of 0.1% (w/v/) SDS was 1.37.
Lot-specific data Generated at ATCC Generated by multiple ARMWG collaborators for each parameter
Fill volume
1.0 mL 0.5 mL
Storage
-70°C or colder -70°C or colder
pH
Not tested 8.0 at room temperature

NIU = NAS infectious units
NAS = normalized adjusted standard
NGS = next generation sequencing
BSA = Bovine serum albumin
ARMWG = Adenovirus Reference Material Working Group
EU = Endotoxin units