Partitioning and inactivation of viruses by the caprylic acid precipitation followed by a terminal pasteurization in the manufacturing process of horse immunoglobulins

Mpandi, M.; Schmutz, P.; Legrand, E.; Duc, R.; Geinoz, J.; Henzelin-Nkubana, C.; Giorgia, S.; Clerc, Olivier; Genoud, D.; Weber, Teresa

doi:10.1016/j.biologicals.2007.02.004

Cited by 20 publications

(11 citation statements)

References 20 publications

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“…Most importantly, manufacturing processes should include one or two dedicated viral inactivation steps, a major tripod of viral safety [30,32]. Caprylic acid treatment is known to be a robust viral reduction treatment for both human [16,18,33] and horse-derived IgG [34,35]. Our study confirms that at the concentration and pH used in this work, this is highly effective against lipid-enveloped viruses, as > 4 log of HIV, BVDV and PRV were inactivated within 15 minutes of treatment.…”

Section: Discussionmentioning

confidence: 99%

Minipool Caprylic Acid Fractionation of Plasma Using Disposable Equipment: A Practical Method to Enhance Immunoglobulin Supply in Developing Countries

El‐Ekiaby¹,

Vargas

Moussa

et al. 2015

PLoS Negl Trop Dis

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BackgroundImmunoglobulin G (IgG) is an essential plasma-derived medicine that is lacking in developing countries. IgG shortages leave immunodeficient patients without treatment, exposing them to devastating recurrent infections from local pathogens. A simple and practical method for producing IgG from normal or convalescent plasma collected in developing countries is needed to provide better, faster access to IgG for patients in need.Methodology/Principal FindingsIgG was purified from 10 consecutive minipools of 20 plasma donations collected in Egypt using single-use equipment. Plasma donations in their collection bags were subjected to 5%-pH5.5 caprylic acid treatment for 90 min at 31°C, and centrifuged to remove the precipitate. Supernatants were pooled, then dialyzed and concentrated using a commercial disposable hemodialyzer. The final preparation was filtered online by gravity, aseptically dispensed into storage transfusion bags, and frozen at <-20°C. The resulting preparation had a mean protein content of 60.5 g/L, 90.2% immunoglobulins, including 83.2% IgG, 12.4% IgA, and 4.4% IgM, and residual albumin. There was fourfold to sixfold enrichment of anti-hepatitis B and anti-rubella antibodies. Analyses of aggregates (<3%), prekallicrein (5-7 IU/mL), plasmin (26.3 mU/mL), thrombin (2.5 mU/mL), thrombin-like activity (0.011 U/g), thrombin generation capacity (< 223 nM), and Factor XI (<0.01 U/mL) activity, Factor XI/XIa antigen (2.4 ng/g) endotoxin (<0.5 EU/mL), and general safety test in rats showed the in vitro safety profile. Viral validation revealed >5 logs reduction of HIV, BVDV, and PRV infectivity in less than 15 min of caprylic acid treatment.Conclusions/Significance90% pure, virally-inactivated immunoglobulins can be prepared from plasma minipools using simple disposable equipment and bag systems. This easy-to-implement process could be used to produce immunoglobulins from local plasma in developing countries to treat immunodeficient patients. It is also relevant for preparing hyperimmune IgG from convalescent plasma during infectious outbreaks such as the current Ebola virus episode.

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Section: Discussionmentioning

confidence: 99%

Minipool Caprylic Acid Fractionation of Plasma Using Disposable Equipment: A Practical Method to Enhance Immunoglobulin Supply in Developing Countries

El‐Ekiaby¹,

Vargas

Moussa

et al. 2015

PLoS Negl Trop Dis

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“…In addition, a modest decrease in the high molecular weight (HMW) aggregates was observed. Finally, OA treatment has also demonstrated efficient inactivation against enveloped viruses and moderately efficient against non‐enveloped viruses (Brodsky, ; Dichtelmuller et al, ; Mpandi et al, ; Vacante and Connell‐Crowley, ). The viral inactivation by OA is attributed to the capability in destroying the lipid bilayer of enveloped viruses and has the potential to aid in viral clearance.…”

Section: Precipitation Technologiesmentioning

confidence: 99%

Clarification technologies for monoclonal antibody manufacturing processes: Current state and future perspectives

Singh

Arunkumar

Chollangi

et al. 2015

Biotech & Bioengineering

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Considerable progress has been made increasing productivity of cell cultures to meet the rapidly growing demand for antibody biopharmaceuticals through increased cell densities and longer culture times. This in turn has dramatically increased the burden of process and product related impurities on the purification processes. In addition, current trends in the biopharmaceutical industry point toward both increased productivity and targeting smaller patient populations for new indications. Taken together, these developments are driving the industry to explore alternative separation technologies as a future manufacturing strategy. Clarification technologies well established in other industries, such as flocculation and precipitation are increasingly considered as a viable solution to address this bottleneck in antibody processes. However, several technical issues need to be fully addressed including suitability as a platform application, robustness, process cost, toxicity, and clearance. This review will focus on recent efforts to incorporate new generation clarification technologies for mammalian cell cultures producing monoclonal antibodies as well as challenges to their implementation supported by a case study.

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“…Caprylic acid has been used to precipitate non-IgG proteins from a solution, leaving an IgG-rich supernatant (Bernard et al, 1996). This technique has been adapted to determine the IgG content of colostrum (Morrill et al, 2012), mammary secretions from nonlactating dairy cattle (Guidry and O'Brien, 1996), and for the purification of serum for therapeutic uses in humans (Perosa et al, 1990;Rojas et al, 1994;Parkkinen et al, 2006;Bergmann-Leitner et al, 2008) and horses (Mpandi et al, 2007). In the neonatal calf, IgG constitutes a large proportion of the protein in serum, suggesting that fractionation with caprylic acid and analysis of supernatants by refractometer could improve estimates of serum IgG concentrations in calves less than 24 h old compared with measuring total protein (TP).…”

Section: Introductionmentioning

confidence: 99%

Estimate of serum immunoglobulin G concentration using refractometry with or without caprylic acid fractionation

Morrill

Polo²,

Lago³

et al. 2013

Journal of Dairy Science

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Objectives of this study were to develop a rapid calf-side test to determine serum IgG concentrations using caprylic acid (CA) fractionation, followed by refractometry of the IgG-rich supernatant and compare the accuracy of this method with results obtained using refractometry using raw serum. Serum samples (n=200) were obtained from 1-d-old calves, frozen (-20°C), and shipped to the laboratory. Samples were allowed to thaw for 1h at room temperature. Fractionation with CA was conducted by adding 1mL of serum to a tube containing 45, 60, or 75µL of CA and 0.5, 1.0, or 1.5mL of 0.06 M acetic acid. The tube contents were mixed well, allowed to react for 1 min, and then centrifuged at 3,300 × g for 0, 10, or 20 min at 25°C. The %Brix and refractive index of the fractionated supernatant were determined using a digital refractometer. Nonfractionated serum was analyzed for %Brix (BRn), refractive index (nDn), and IgG concentration by radial immunodiffusion. The mean serum IgG concentration was 19.0 mg/mL [standard deviation (SD)=9.7], with a range of 3.5 to 47.0 mg/mL. The mean serum BRn was 8.6 (SD=0.91), with a range of 6.8 to 11.0. The mean serum nDn was 1.34566 (SD=0.00140), with a range of 1.34300 to 1.34930. Serum nDn was positively correlated with IgG concentration (correlation coefficient=0.86; n=185). Fractionated samples treated with 1mL 0.6 M acetic acid and 60µL of CA and not centrifuged before analysis resulted in a strong relationship between the refractive index of the fractionated supernatant and IgG (correlation coefficient=0.80; n=45). Regression was used to determine cut points indicative of 10, 12, and 14 mg of IgG/mL to determine the sensitivity and specificity of refractometry to identify failure of passive transfer (serum IgG <10 mg/mL at 24 h old). The nDn were 1.34414, 1.34448, and 1.34480 to predict 10, 12, and 14 mg of IgG/mL of serum, respectively. The BRn cut points were 7.6, 7.8, and 8.0, respectively. The nDn cut points of 1.34448 and 1.34480 resulted in similar specificities (82.9%), whereas the 1.34414 cut point had a specificity of 60.0%. The BRn cut point of 7.6 and 7.8%Brix resulted in a similar percentage of correctly classified samples (89.7 and 90.8%, respectively); however, the 7.8% Brix cut point resulted in fewer false positives. These results suggest that Brix refractometry of nonfractionated calf serum provides a strong estimate of IgG concentration and 7.8% Brix may be used as the cut point to identify failure of passive transfer in 1-d-old calves.

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Partitioning and inactivation of viruses by the caprylic acid precipitation followed by a terminal pasteurization in the manufacturing process of horse immunoglobulins

Cited by 20 publications

References 20 publications

Minipool Caprylic Acid Fractionation of Plasma Using Disposable Equipment: A Practical Method to Enhance Immunoglobulin Supply in Developing Countries

Minipool Caprylic Acid Fractionation of Plasma Using Disposable Equipment: A Practical Method to Enhance Immunoglobulin Supply in Developing Countries

Clarification technologies for monoclonal antibody manufacturing processes: Current state and future perspectives

Estimate of serum immunoglobulin G concentration using refractometry with or without caprylic acid fractionation

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