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N.C.A Lu177 Reference Radiolabeling Procedure

The document provides instructions for radiolabeling PSMA and DOTA peptides with the radionuclide lutetium-177. Lutetium-177 is manufactured by Eczacıbaşı Monrol Nükleer Ürünler A.Ş under cGMP conditions and allows obtaining therapeutic doses of radiolabeled peptides with high specific activity and radionuclide purity. The document outlines the requirements, preparation, radiolabeling procedure, quality control testing using HPLC or TLC, and an example radiolabeling procedure for 200 mCi of lutetium-177.

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Fadhil Muhammad A.
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130 views

N.C.A Lu177 Reference Radiolabeling Procedure

The document provides instructions for radiolabeling PSMA and DOTA peptides with the radionuclide lutetium-177. Lutetium-177 is manufactured by Eczacıbaşı Monrol Nükleer Ürünler A.Ş under cGMP conditions and allows obtaining therapeutic doses of radiolabeled peptides with high specific activity and radionuclide purity. The document outlines the requirements, preparation, radiolabeling procedure, quality control testing using HPLC or TLC, and an example radiolabeling procedure for 200 mCi of lutetium-177.

Uploaded by

Fadhil Muhammad A.
Copyright
© © All Rights Reserved
Available Formats
Download as PDF, TXT or read online on Scribd
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n.c.a.

LUTETIUM-177
manufactured by Eczacıbaşı Monrol Nükleer Ürünler A.Ş.

Reference radiolabeling procedure for PSMA & DOTA with n.c.a. 177Lu in 200-600 µl 0.05 M HCl

177Lu for Radiolabeling

Eczacıbaşı Monrol manufactures non-carrier-added 177Lu under cGMP conditions. It allows obtaining
therapeutic doses of radiolabeled 177Lu-PSMA & 177Lu-DOTA with high specific activity and radionuclide
purity.

Non-carrier-added 177Lu is not intended for use in humans. It has not been
registered as a pharmaceutical product.

Preparation of 177Lu-PSMA & 177Lu-DOTA


Requirements
1. Sodium L-Ascorbate (Ultra pure)
2. L-Ascorbic Acid (Ultra pure)
3. Water (Ultra pure)
4. PSMA 617 / I&T
5. DOTA-TATE / TOC / NOC
6. 0.9 % Saline
7. 0.22 µm Sterile filter
8. Sterile Vial
9. Micro-pipettes with tips
10. pH Strips with range (4-6) or Electronic pH meter for micro fluids
11. Sterican needles
12. Digital hot pot or Fully automated synthesizer
13. QC accessories (As per user standards/practices)

Peptide Amount Calculation (PSMA & DOTA)


- PSMA; 0.5 µg per 1 mCi 177Lu
- DOTA; 0.65 µg per 1 mCi 177Lu

Buffer Preparation
- 80 mg Sodium L-ascorbate is dissolved in 1 ml ultra pure water.
- 20 mg L-ascorbic acid is dissolved in 1 ml ultra pure water.
- Mix the whole mixture in a vial & dissolve completely with shaking.
- Buffer is ready to use.

Radiolabelling
- Add calculated volume of peptide to buffer vial.
- Mix well, check pH value (pH 4-6).
- Add the above buffer to the Lutetium vial with the help of Sterican needle.
- Heat the vial at 100°C for 25 minutes.
- Allow it to cool for 5 minutes & then put a vent.
* 177Lu bound peptide compound can optionally be purified at this step with the C18 Light
cartridge.

C18 Light Cartridge Preconditioning


Preconditioning with 5 mL Ethanol, purged with air to dryness, 10 mL Inj. Water, purged
with air to dryness)

The 177Lu bound peptide compound is passed through the preconditioned C18 Light
cartridge. The labeled product remains in the C18 Light cartridge and unbound 177Lu
goes to waste then 2 ml 50% EtOH-Inj.water solution passes through C18 Light cartridge
and carries bound lutetium to the final product vial.

- Adjust the volume to 10 ml with 0.9 % saline.


- Pass the solution to 0.22 µm sterile filter.
- Do necessary QC.
Radiolabeling example:

for 200 mCi 177Lu in 200 µl 0.05 M HCl

- 130 µg DOTA is dissolved in 130 µl ultra pure water.


- 80 mg Sodium L-ascorbate is dissolved in 1 ml ultra pure water, transferred to the DOTA vial
(washing the vial walls).
- 20 mg L-ascorbic acid is dissolved in 1 ml ultra pure water, transferred to the DOTA vial (washing
the vial walls).
- Mix well, check pH value (pH 4-6).
- Add the peptide and buffer mixture to the Lutetium vial with the help of a sterican needle.
- The reaction mixture is heated for 25 minutes at 100°C. The solution remains clear and
light yellow.
- Allow it cool, 5 to 20 µl aliquot is used for QC (either TLC or HPLC).
- For the application the reaction mixture is adjusted to 10 ml with isotonic sodium
chloride solution and underwent the sterile filtration.

Quality Control

Requirements for HPLC


1. HPLC Column; C18 5 µm, 4.6 x 250 mm
2. Trifluoroacetic acid (Ultra pure)
3. Acetonitrile (HPLC grade)
4. Water (Ultra pure)

HPLC Method

Column
Size: l = 0.25 m, Ø = 4.6 mm
Stationary phase: Reversed phase Columns (C18 Column) for chromatography R (5 µm)
Mobile phase
mobile phase A: trifluoroacetic acid R,water R water for chromatography R (1:999 V/V)
mobile phase B: trifluoroacetic acid R, acetonitrile R (1:999 V/V)
Wavelength
220 nm
Gradient for PSMA
Flow Rate: 1 ml/min
0-8 min 0% B; 8-12 min 0-75% B, 12-12.01 min 75% B, 12.01-16 min 0% B
Gradient for DOTA
Flow Rate: 1.3 ml/min
0-8 min 24% B; 8-9 min 24-60% B, 9-14 min 60% B, 14-14.01 min 60-24% B, 14.01-20 min 24% B

Requirements for TLC


1. Ammonium Acetate (Ultra pure)
2. Methyl Alcohol 99.9%
3. Water (Ultra pure)
4. TLC Strips & Chromatography chamber
TLC Method

Solvent Preparation for TLC

1 M Ammonium Acetate 50:50 Methyl Alcohol (Solvent for Chromatography)

- Weigh 7.7 g of Ammonium Acetate.


- Take 100 mL of ultra pure water in measuring cylinder & transfer in the conical flask.
- Mix the weighed 7.7 g Ammonium Acetate to measured water.
- Add 100 ml of Methyl Alcohol to the above flask & mix well.
- Your solvent is ready to use.

Procedure
- Cut ITLC-SG strips of size 7.0 cm X 2.0 cm.
- Pour about 7-8 mm of QC solvent (1 M Ammonium Acetate 50:50 Methyl Alcohol) to
chromatography chamber.
- Cover the chamber with glass plate & leave it for 10 minutes.
- Mark the ITLC strips with aliquot 1 cm above from bottom edge.
- After drying transfer the strip to chromatography chamber & cover it with glass sheet.
- Let the solvent to rise till app. 6 cm. from bottom.
- Labelled product will be migrated with the solvent front (Rf = 0.9 - 1) & on contrary Free
Lu will remain at origin.

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