(M0930-07-41) Use of 5% Dextrose Intravenous Diluent for Therapeutic Monoclonal Antibodies: Impact on Mabs Under Varying Formulation Conditions

Purpose: 5% Dextrose (D5W) is commonly used as the alternative IV diluent to 0.9% sodium chloride (saline)¹, enabling flexibility for healthcare providers and providing clinical benefits over high salt saline for patients with hypernatremia² and renal insufficiency^3,4. Loss of stability can occur when a protein formulation is diluted to low concentrations with IV solutions due to many factors such as protein properties, formulation composition, concentration of the active pharmaceutical ingredient, choice of diluent, and product contact surface.⁵ Glycation (non-enzymatic glycosylation) has also been reported after dilution with dextrose for some mAb molecules, because dextrose as a type of reducing sugar is known to react with the susceptible amine group (primarily on the lysine side chain) to form Schiff base intermediate and stable ketoamine upon Amadori rearrangement. ⁶ In addition, environmental effects can also result in incompatibility of mAbs diluted in IV diluents because of physical and temperature stress during handling such as transportation or storage at ambient temperature, mixing, and agitation.⁷ This study examines the influence of formulation composition on the physico-chemical in-use stability of distinct monoclonal antibody (mAb) formats when using 5% dextrose (D5W) for intravenous administration under clinical in-use conditions.

Methods: Four distinct mAbs of different frameworks were studied to evaluate the impact of various formulation parameters (including pH, ionic strength, and buffer types) on physico-chemical in-use stability according to Table 1. A scaled down model in polyethylene terephthalate glycol (PETG) bottles was used to mimic the IV bag throughout this study. The 30mg/mL mAb drug products in a given formulation were diluted into the respective IV diluents (D5W or 0.9% sodium chloride) to achieve a 1.5mg/mL protein concentration.  The diluted samples were stored at 30°C for 24 hours, representing the worst-case ambient temperature in clinics, and then subjected to analytical testing for product quality assessments. The physico-chemical stability of therapeutic proteins was evaluated with visual inspection for visible particulates, UV spectroscopy for turbidity, Size-Exclusion Chromatography (SEC) for aggregates, Ion-Exchange Chromatography (IEC) for charge isoforms and Liquid chromatography–mass spectrometry (LC-MS) for glycation analysis.

Results: For physical in-use stability, little or no differences were observed on % aggregation by SEC for all four mAbs once diluted in IV diluent (D5W or saline control) across various formulation conditions (F1 to F14).  For chemical in-use stability, all four mAbs diluted in D5W resulted in an elevated level of acidic variants relative to its respective saline control across F1 to F14 (at T0 as well as post 24 hours at 30°C). In addition, pH increase from 5.0 (F1) to 8.0 (F5) resulted in an increasing trend of glycation and acidic variant formation for all four mAbs in D5W (Figure 1). It also exhibited a strong-positive correlation in between acidic charge variant and total glycation formation (Figure 2). These results demonstrated a positive impact of formulating the four mAbs at acidic pH with less or minimal impact on chemical stability including both charge variant and glycation formation when diluted in D5W. However, similar levels of acidic variants for mAb 1-4 were observed after dilution in D5W at T0 as well as post 24 hours at 30°C, regardless of the ionic strength or the buffer system employed (F6-F14).

Conclusion: The in-use stability results showed that chemical stability is the most sensitive stability indicator for compatibility assessment of D5W diluents with no significant impact on physical stability across various formulation parameters. A strong-positive correlation was established in between acidic charge variant and total glycation formation, suggesting HPLC-based charge assays may serve as a high throughput screening method for D5W in-use stability assessment. Within the dataset evaluated, neither the ionic strength nor buffer types played a role on mAb in-use stability with D5W. These results and observations contribute to understanding the impact of formulation parameters on the use of D5W as a diluent for IV infusion administration, providing valuable insights for the future pharmaceutical development of therapeutic proteins.

References: 1 Voldby AW, Brandstrup B 2016. Fluid therapy in the perioperative setting-a clinical review. J Intensive Care 4:27.
2 Tauseef A, Zafar M, Syed E, Thirumalareddy J, Sood A, Lateef N, Mirza M 2021. Prognostic importance of deranged sodium level in critically ill patients: A systemic literature to review. J Family Med Prim Care 10(7):2477-2481.
3 Ding X, Cheng Z, Qian Q 2017. Intravenous Fluids and Acute Kidney Injury. Blood Purif 43(1-3):163-172.
4 Lippold C, Patel A 2020. Correction of hyponatremia by infusing D5W (5% dextrose in water solution) prefilter in patients receiving continuous renal replacement therapy: A case series. Hemodial Int 24(2):E27-E32.
5 Morar-Mitrica S, Puri M, Beumer Sassi A, Fuller J, Hu P, Crotts G, Nesta D 2015. Development of a stable low-dose aglycosylated antibody formulation to minimize protein loss during intravenous administration. MAbs 7(4):792-803.
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7 Sreedhara A, Glover ZK, Piros N, Xiao N, Patel A, Kabakoff B 2012. Stability of IgG1 monoclonal antibodies in intravenous infusion bags under clinical in-use conditions. J Pharm Sci 101(1):21-30.

Acknowledgements: The authors thank Y Diana Liu and Feng Yang for suggestions on differential glycation measurement using LC/MS.


Table 1. Formulation Screening Study Overview


Figure 1. Impact of pH on Δ%glycation relative to DP (Red for %total glycation and Blue for %stable glycation) and Δ%Acidic Regions relative to DP (Black) for mAb1-4 diluted in D5W followed by 24h at 30 °C. (Error bar = one standard deviation, n=3).


Figure 2. Correlation of Δ% total glycation (T24h at 30 °C relative to DP) and Δ%Acidic regions by IEC (T24h at 30 °C relative to starting DP) in D5W for mAb1 – 4 as a function of pH (F1-F5). (Error bar = one standard). deviation, n=3).