Technetium 99mTc Fanolesomab Injection
» Technetium 99mTc Fanolesomab Injection is a sterile, nonpyrogenic preparation of anti-CD15 antibody, a partially reduced murine IgM monoclonal antibody that is labeled with 99mTc and is suitable for intravenous administration. It may contain reducing agents, buffers, and stabilizers. It contains no antimicrobial agents. Other chemical forms of radioactivity do not exceed 10 percent of the total radioactivity.
Caution—Components of the commercial kit that are used to prepare the Injection are not to be administered directly to the patient.
Packaging and storage—
Preserve in single-dose containers. Store in a refrigerator. [note—After radiolabeling, the Technetium Tc 99m Fanolesomab Injection can be kept at room temperature.]
Labeling—
Label it to include the following, in addition to the information specified for Labeling under Injections 
1
: the time and date of calibration; the amount of 99mTc as labeled fanolesomab expressed in MBq (or mCi) per mL at the time of calibration; the expiration date and time; the storage temperature; and the statement “Caution—Radioactive Material”. The labeling indicates that the radioactive half-life of 99mTc is 6.0 hours and that, in making dosage calculations, correction is to be made for radioactive decay. The labeling also states that the Injection is to be used within 6 hours following constitution.
1: the time and date of calibration; the amount of 99mTc as labeled fanolesomab expressed in MBq (or mCi) per mL at the time of calibration; the expiration date and time; the storage temperature; and the statement “Caution—Radioactive Material”. The labeling indicates that the radioactive half-life of 99mTc is 6.0 hours and that, in making dosage calculations, correction is to be made for radioactive decay. The labeling also states that the Injection is to be used within 6 hours following constitution.
Bacterial endotoxins 85:
not more than 0.1 Endotoxin Unit per µg of fanolesomeb in the prepared Injection.
85:
not more than 0.1 Endotoxin Unit per µg of fanolesomeb in the prepared Injection.
pH 791:
between 5.8 and 6.6.
791:
between 5.8 and 6.6.
Particulate matter 788—
It meets the requirements for particulate matter specified for small-volume injections.
788—
It meets the requirements for particulate matter specified for small-volume injections.
Radiochemical purity—
system 1 (Free pertechnetate 99mTc)—
Test solution—
Use the Injection.
Application volume—
Apply 3 µL of the Test solution to the origin.
Developing solvent system:
methyl ethyl ketone (MEK).
Procedure—
Apply the Test solution about 2 cm from the bottom of the Adsorbent strip. Immediately develop by ascending chromatography (see Thin-Layer Chromatography under Chromatography 621) until the solvent front has moved about 7.5 cm from the origin. The radiochemical impurity, free pertechnetate 99mTc, migrates to the top section, while colloidal 99mTc and technetium 99mTc fanolesomab remains near the origin on the bottom section. Allow the chromatogram to air-dry. Determine the distribution of radioactivity on the chromatogram by cutting the developed strip at 4 cm from the bottom and then separately measuring and recording the net radioactivity in the top and bottom sections using a suitable radiation detector. Calculate the percentage of free pertechnetate 99mTc in the Injection by the formula:
621) until the solvent front has moved about 7.5 cm from the origin. The radiochemical impurity, free pertechnetate 99mTc, migrates to the top section, while colloidal 99mTc and technetium 99mTc fanolesomab remains near the origin on the bottom section. Allow the chromatogram to air-dry. Determine the distribution of radioactivity on the chromatogram by cutting the developed strip at 4 cm from the bottom and then separately measuring and recording the net radioactivity in the top and bottom sections using a suitable radiation detector. Calculate the percentage of free pertechnetate 99mTc in the Injection by the formula:
100 AT / (AT + AB)
in which AT is the radioactivity measured on the top section; and AB is the radioactivity measured on the bottom section.
system 2 (Colloidal 99mTc)—
Adsorbent—
Use affinity thin-layer chromatography (ATLC) strips2 that have been soaked in a solution of 50% newborn calf serum (NBCS)3 in water and allowed to air-dry overnight.
Test solution—
Use the Injection.
Developing solvent system:
4% alcohol in 0.3 M sodium chloride.
Application volume—
Prespot the point of origin with 3 µL of Developing solvent system followed immediately by 3 µL of the Test solution.
Procedure—
Apply the Test solution about 2 cm from the bottom of the Adsorbent strip. Immediately develop the strip by ascending chromatography (see Thin-Layer Chromatography under Chromatography 621) until the solvent front has moved about 7.5 cm above the origin. The radiochemical impurity, colloidal 99mTc, will remain at the origin, while free pertechnetate 99mTc and technetium 99mTc fanolesomab migrate close to the solvent front. Remove the strip, and allow to air-dry. Cut the strip 4 cm from the bottom, and separately measure and record the background-corrected radioactivity found in the top and bottom sections, using a suitable radiation detector. Calculate the percentage of colloidal 99mTc by the formula:
621) until the solvent front has moved about 7.5 cm above the origin. The radiochemical impurity, colloidal 99mTc, will remain at the origin, while free pertechnetate 99mTc and technetium 99mTc fanolesomab migrate close to the solvent front. Remove the strip, and allow to air-dry. Cut the strip 4 cm from the bottom, and separately measure and record the background-corrected radioactivity found in the top and bottom sections, using a suitable radiation detector. Calculate the percentage of colloidal 99mTc by the formula:
100 AB / (AB + AT)
in which AB is the radioactivity measured in the bottom section; and AT is the radioactivity measured in the top section. The sum of the result for free pertechnetate 99mTc in System 1 and for colloidal 99mTc measured in System 2 is not more than 10%.
Immunoreactivity—
Adsorbent—
Use affinity thin-layer chromatography (ATLC) strips2 that have been soaked in a solution of 50% newborn calf serum (NBCS)3 in water and allowed to air-dry overnight.
Diluent—
Prepare a mixture of NBCS and pH 7.4 phosphate-buffered saline (PBS) (1:1).
Test solution—
Use the Injection diluted with Diluent (1 in 4).
reference materials—
Positive control:
CD15 positive HL-60 (ATCC No. CCL240) formalin fixed-cell suspension (12 × 106 cells per mL).
Negative control:
CD15 negative Raji (ATCC No. CCL86) formalin fixed-cell suspension (12 × 106 cells per mL).
Application volume—
Thaw, and mix the HL-60 and Raji cell stock suspensions. Dispense 90-µL aliquots into individual incubation tubes. Add 3 µL of the Test solution to each incubation tube, and mix on a vortex mixer for 5 seconds. Incubate for 30 minutes with gentle rocking at 37 ± 2
.
.
Developing solvent system—
Prepare a solution containing 0.05% sodium azide and 4% alcohol in 10 mM phosphate-buffered saline, pH 7.4 (PBS). Pass through a filter having a 0.22-µm porosity.
Procedure—
Mix each incubation tube on a vortex mixer. Immediately remove, and apply 10 µL of sample to the origin of the Adsorbent ATLC strip (2 cm from bottom). Allow the sample to adsorb onto the strip, and immediately develop the strip by ascending chromatography (see Thin-Layer Chromatography under Chromatography 621) until the solvent front has moved about 7.5 cm from the origin. Remove the strips, and allow to air-dry. Cut both strips 4 cm above the origin. Separately measure and record the background-corrected radioactivity found on the top and bottom sections of each strip, using a suitable radiation detector. Immunoreactive technetium 99mTc fanolesomab, bound to the HL-60 cells, remains at the origin, while nonbound forms of 99mTc migrate away from the origin. Nonspecific binding is measured using the Raji negative control cells. Calculate the specific immunoreactive binding by the formula:
621) until the solvent front has moved about 7.5 cm from the origin. Remove the strips, and allow to air-dry. Cut both strips 4 cm above the origin. Separately measure and record the background-corrected radioactivity found on the top and bottom sections of each strip, using a suitable radiation detector. Immunoreactive technetium 99mTc fanolesomab, bound to the HL-60 cells, remains at the origin, while nonbound forms of 99mTc migrate away from the origin. Nonspecific binding is measured using the Raji negative control cells. Calculate the specific immunoreactive binding by the formula:
[100AB(HL-60) /(AB(HL-60) + AT(HL-60))] – [100AB(Raji) /(AB(Raji) + AT(Raji))]
in which AB(HL-60) and AB(Raji) are the radioactivity of the HL-60 positive control cells and Raji negative control cells, respectively, measured on the bottom section of each strip; and AT(HL-60) and AT(Raji) are the radioactivity of the HL-60 positive control cells and Raji negative control cells measured on the top section of each strip. A minimum specific immunoreactive binding of 40% is required for the CD15-positive HL-60 cells.
Other requirements—
It meets the requirements for Radionuclide identification and Radionuclidic purity under Sodium Pertechnetate Tc 99m Injection. It also meets the requirements under Injections 1, except that it may be distributed or dispensed prior to completion of the test for Sterility, the latter test being started on the date of manufacture.
1, except that it may be distributed or dispensed prior to completion of the test for Sterility, the latter test being started on the date of manufacture.
Assay for radioactivity 821—
Using a suitable counting assembly (see Selection of a Counting Assembly), determine the radioactivity, in MBq (or mCi) per mL, of Injection by use of a calibrated system.
821—
Using a suitable counting assembly (see Selection of a Counting Assembly), determine the radioactivity, in MBq (or mCi) per mL, of Injection by use of a calibrated system.
1
ITLC strips for free pertechnetate 99mTc may be obtained from Sunset Scientific Strips LLC (product number 10503), P.O. Box: 3895, Albuquerque, NM 87190-3895.
2
ATLC strips for colloidal 99mTc may be obtained from Sunset Scientific Strips LLC (product number 10506), P.O. Box: 3895, Albuquerque, NM 87190-3895.
3
NBCS (heat inactivated) may be obtained from GIBCO/Invitrogen Corp. (catalog number 26010-074), 1600 Faraday Avenue, P.O. Box 6482, Carlsbad, CA 92008
Auxiliary Information—
Please check for your question in the FAQs before contacting USP.
| Topic/Question | Contact | Expert Committee |
| Monograph | Ian DeVeau, Ph.D.
Director, Veterinary Drugs and Radiopharmaceuticals 1-301-816-8178 |
(RMI05) Radiopharmaceuticals and Medical Imaging Agents 05 |
| Reference Standards | Lili Wang, Technical Services Scientist 1-301-816-8129 RSTech@usp.org |
|
| 85 |
Radhakrishna S Tirumalai, Ph.D.
Senior Scientist 1-301-816-8339 |
(MSA05) Microbiology and Sterility Assurance |
USP32–NF27 Page 3662
Pharmacopeial Forum: Volume No. 31(5) Page 1405