[68Ga] peptide high-output production on commercially

available MiniAIO® synthesizer

Objectives

Optmization parameters

Material and Methods

References

Conclusion/Perspective

Adapt reaction media pH, temperature, durationsensitivity of biomolecules

Development of an easy high-output Gallium-68 peptide labeling process1

Collet C.1,2, Remy S. 1,2, Didier R1,2.,Vergote T.4, Véran N. 3, Karcher G.1,2,3

1-Université de Lorraine, F-54500 Vandoeuvre les Nancy – France;

2-NancycloTEP, Plateforme d’imagerie moléculaire, 54500 Vandoeuvre les Nancy – France;

3-CHRU de Nancy-Brabois, F-54511 Vandoeuvre les Nancy – France

4-TRASIS SA, B-4430 Ans-Belgium

Elution

Peptide labelling

Delivery

Filtre integrity test

Concept performed on commercially available peptides

[1] Breeman W, de Blois E, Chan HS, et al. [2011], Semin Nucl Med, 41: 314-321

[2] Velikyan [2014], Theranostic, 4(1): 47-80

Peptide formulated in 1 mL of a specific buffer

Generator eluted with 5 mL of HCl 0.1 M as recommended by the supplier

AcONa 1,5 M gives better results

Specific buffer (neutralisant media)

• Temperature influence

• Reaction times influence

N

N

NNpeptide

N

N

NN

peptide68Ga

IGG100Low germanium

breakthrough

Commercial

peptideGMP or/and not GMP

mAIO synthetiser

from TrasisSingle use cassette

Methods without pre and post purification

Reproducible and robust

methods to label peptide

with Gallium-68

Constant parameters

Peptide precursor quantity

influence

Quality Control

Constant parameters: AcONa 1.5 M; 10 min;

NODAGA-RGD and PSMA-11 30°C; DOTATOC and DOTANOC 95°C

Constant parameters: AcONa 1.5 M; 10 min;

NODAGA-RGD and PSMA-11 5µg; DOTATOC,DOTANOC 30µg

Constant parameters: AcONa 1,5 M; NODAGA-RGD and

PSMA-11 30°C; DOTATOC and DOTANOC 95°C; NODAGA-

RGD and PSMA-11 5µg; DOTATOC and DOTANOC 30µg

NODAGA-

RGD

PSMA-11 DOTATOC DOTANOC

Neutralisant AcONa

1,5 M

AcONa

1,5 M

AcONa

1,5 M

AcONa

1,5 M

Precursor

quantity

5 µg 5 µg 30 µg 30 µg

Duration 10 min 10 min 10 min 10 min

Temperature 30°C 30°C 95°C 95°C

RCY* (dc) 95% 96% 96% 95%

Synthesis

duration

15 min 15 min 15 min 15 min

BEST CONDITION

• HPLC : UV and radio detection

Analysis condition: 0% 2 min, Gradient 0 to 40 % in 14 min

Column ACE C18 flow 0,5 mL/min; injection 20µL

Detection at 220 nm

[69Ga]NODAGA-RGD : LOD = 0.6 µg/mL; LOQ = 1.8 µg/mL

NODAGA-RGD : LOD = 0.4 µg/mL; LOQ = 1.3 µg/mL

• Validation Batches

Test Nominal ValueBatch n° 1 Batch n° 2 Bacht n° 3

Results C Results C Result C

pH value 4,0-8,0 7 YES 7 YES 7 YES

Appearence Clear, colourless solutionClear, colourless

solutionYES

Clear, colourless

solutionYES

Clear, colourless

solutionYES

Radiochemical purity ≥ 91% >98% YES >98% YES >98% YES

Radionuclidic purity > 99% >99% YES >99% YES >99% YES

Radionuclear identity Half life 62 -74 min 68 min YES 66 min YES 67 min YES

Sterility Conforme Ph.Eur. Sterile YES Sterile YES Sterile YES

Filter integrity Trasis specification > 3 bars YES > 3 bars YES > 3 bars YES

One representative exemple is done with NODAGA-RGD

Simple and reproductible synthesis methods without solvent

Compatible with different peptide precursors

Excellent radiochemical yield

Each radiotracer was synthesized more than 10 times

Germanium concentration lower than the Ph.Eur. limit

Each radiotracors [68Ga]NODAGA-RGD, [68Ga]PSMA-11, [68Ga]DOTATOC and

[68Ga]DOTANOC have this specification.

NH

O

HN

NH

O

OHN

O

HN

O

HN

NH

HN

HO NH2

O

O

O

NH

HOHO

S

S

NN

N N

OHO O O

OO

68Ga3+

NH

O

HN

NH

O

OHN

O

HN

O

HN

NH

HN

HO NH2

O

O

O

NH

HOHO

S

S

NN

N N

OHO O O

OO

68Ga3+OH

HN

HN

NH

NH

NH

O

O

O

O

O

HN

COO

O

HOOC

N

N

N

COO

COO

NH

NH2

NH

68Ga3+

TyrLys

N

68Ga3+O O

N

O

O

O

O

COOH

O

HN

NH

O

NH

COOHNH

O

HOOC

COOH

[69Ga]NODAGA-RGD

NODAGA-RGD

[69Ga]DOTATOC

DOTATOC

Waters e2695

[69Ga]PSMA-11

PSMA-11

[69Ga]DOTANOC

DOTANOC

Analysis condition: 24% 8 min, Gradient 24 to 60 % in 1 min

Column ACE C18 flow 0,6 mL/min; injection 20µL

Detection at 220 nm

[69Ga]DOTATOC : LOD = 0.7 µg/mL; LOQ = 2.1 µg/mL

DOTATOC : LOD = 0.8µg/mL; LOQ = 2.35 µg/mL

Analysis condition: Gradient 0 to 75 % in 8 min, 75% in 4 min

Column Waters Xterra RPC18 flow 1mL/min; injection 20µL

Detection at 220 nm

Analysis condition: 30% 2 min, Gradient 30 to 80 % in 15 min

Colonne ACE C18 flow 0,5 mL/min; injection 20µL

Detection at 220 nm

[69Ga]DOTANOC : LOD = 0.6 µg/mL; LOQ = 1.7 µg/mL

DOTANOC : LOD = 0.7 µg/mL; LOQ = 2.2 µg/mL[69Ga]PSMA-11 : LOD = 1.8 µg/mL; LOQ = 5.6 µg/mL

PSMA-11 : LOD = 2.0 µg/mL; LOQ = 6 µg/mL

[69Ga]DOTANOC

DOTANOC

HPLC methods are specific, repeatable, robust, reproductible and linear

[68Ga]DOTANOC

[68Ga]DOTATOC

[68Ga]PSMA-11[68Ga]NODAGA-RGD

No organic solvent used during the synthesis for a QC without Gaz Chromatography