Journal: Pathology International

Article Title: Neonatal Fc receptor induces intravenous immunoglobulin growth suppression in Langerhans cell histiocytosis

doi: 10.1111/pin.13068

Figure Lengend Snippet: Neonatal Fc receptor (FcRn) knockdown abrogates intravenous immunoglobulin therapy (IVIG) preparation‐induced growth suppression of ELD‐1 cells in RPMI1640 supplemented with albumin, but not in RPMI1640 supplemented with glutamine or in RPMI1640 alone. ( a ) FcRn knockdown in ELD‐1 cells. Immunoblotting and CCK‐8 assay. Mock or FcRn‐knockdown ELD‐1 cells were incubated for 12 h with or without IVIG preparation in ( b ) RPMI1640 only or ( c ) RPMI1640 supplemented with albumin (n = 3, respectively). Growth was assessed as described in the Materials and Methods. Relative values are compared to the growth of mock ELD‐1 cells without IVIG preparation, which were set to 100.

Article Snippet: Mock or FcRn‐knockdown ELD‐1 cells were cultured for 46 h at 1–3 × 10 4 cells/100 μL of RPMI1640 only (glutamine‐free and albumin‐free) or RPMI1640 supplemented with albumin, with or without IVIG preparation (15 mg/mL, Privigen, CSL Behring AG, Bern, Switzerland) which was dialyzed against large volumes of RPMI1640 only at 4°C using Slide‐A‐Lyzer γ‐irradiated dialysis cassettes (Pierce, Rockford, IL)., We added 10 μL of CCK‐8 solution for the last 2 h and estimated the absorbance at 450 nm, according to the manufacturer's instructions.

Techniques: Knockdown, Western Blot, CCK-8 Assay, Incubation

Journal: Pathology International

Article Title: Neonatal Fc receptor induces intravenous immunoglobulin growth suppression in Langerhans cell histiocytosis

doi: 10.1111/pin.13068

Figure Lengend Snippet: Neonatal Fc receptor (FcRn) enhances intravenous immunoglobulin therapy (IVIG) preparation‐induced recycling of albumin and suppression of albumin‐dependent cell growth in ELD‐1 cells. ( a ) Residual fluorescein isothiocyanate (FITC)‐conjugated albumin in the supernatant of ELD‐1 cells. Mock or FcRn‐knockdown ELD‐1 cells were incubated for 48 h with or without IVIG preparation in RPMI1640 with 0.1% albumin (n = 3, respectively). The cell free supernatants were collected and their absorbance was measured at 490 nm. Relative values are indicated by the absorbance without IVIG preparation, which was set to 100. * P < 0.05 compared with no treatment. IVIG preparation‐induced ELD‐1 cells containing albumin in a FcRn‐dependent manner. Mock or FcRn‐knockdown ELD‐1 cells were incubated for 0 or 2 h with IVIG preparation in RPMI1640 with 0.1% albumin. The cells were collected, washed, and analyzed by ( b ) immunoblotting. Representative data is shown. ( c ) FcRn is required for localization with albumin in Rab11‐positive recycle endosomes in ELD‐1 cells, and IVIG preparation treatment prolongs this process. Mock or FcRn‐knockdown ELD‐1 cells were treated with 0.1% FITC‐conjugated albumin with or without 15 mg/mL IVIG preparation for 0, 30 or 120 min. Cells were fixed in 4% (v/v) paraformaldehyde and stained with anti‐Rab11 antibody, followed by goat anti‐mouse IgG H&L (Alexa Fluor 488). The slides were mounted with fluoroshield mounting medium with DAPI. The closed arrowheads indicate colocalization of Rab11 and albumin.

Article Snippet: Mock or FcRn‐knockdown ELD‐1 cells were cultured for 46 h at 1–3 × 10 4 cells/100 μL of RPMI1640 only (glutamine‐free and albumin‐free) or RPMI1640 supplemented with albumin, with or without IVIG preparation (15 mg/mL, Privigen, CSL Behring AG, Bern, Switzerland) which was dialyzed against large volumes of RPMI1640 only at 4°C using Slide‐A‐Lyzer γ‐irradiated dialysis cassettes (Pierce, Rockford, IL)., We added 10 μL of CCK‐8 solution for the last 2 h and estimated the absorbance at 450 nm, according to the manufacturer's instructions.

Techniques: Knockdown, Incubation, Western Blot, Staining

Journal: Transfusion Medicine and Hemotherapy

Article Title: Effect of Methylene Blue Pathogen Inactivation on the Integrity of Immunoglobulin M and G

doi: 10.1159/000514485

Figure Lengend Snippet: IgG Fc binding to THP-1 cell Fc receptors. a IVIG was used as positive control, binding analysis was feasible with concentrations above 0.1 µg/mL (reference for plasma concentrations 7–16 × 103 µg/mL; n = 1). b IgG binding to Fc receptors on THP-1 cells did not differ before and after MB pathogen inactivation (n = 10 individual donor plasma units, means ± SD are given). ns, not significant.

Article Snippet: IVIG (Privigen®, CSL Behring GmbH, Hattersheim am Main, Germany) was used as positive control and buffer as negative control, as previously described [ 14 ].

Techniques: Binding Assay, Positive Control

Journal: Biodrugs

Article Title: Reduction of Isoagglutinin in Intravenous Immunoglobulin (IVIG) Using Blood Group A- and B-Specific Immunoaffinity Chromatography: Industry-Scale Assessment

doi: 10.1007/s40259-016-0192-3

Figure Lengend Snippet: Original and modified processes for the manufacture of the intravenous immunoglobulin product, Privigen. a When assessing the clearance of immunoglobulin M isoagglutinins by the anion-exchange chromatography and immunoaffinity chromatography steps, the process steps outlined in orange (ethanol- and octanoic acid fractionation) were omitted to minimize the loss of immunoglobulin M, and therefore the loss of the immunoglobulin M isoagglutinins. This was done because the octanoic acid fractionation step is the main elimination step of total immunoglobulin M. The intention was to retain as much immunoglobulin M as possible for the subsequent isoagglutinin removal steps. AIC immunoaffinity chromatography, AIEX anion exchange chromatography, DF diafiltration, UF ultrafitration. Adapted with permission from Hoefferer et al. © 2015 AABB. Hoefferer L, Glauser I, Gaida A, Willimann K, Marques Antunes A, Siani B, et al. Isoagglutinin reduction by a dedicated immunoaffinity chromatography step in the manufacturing process of human immunoglobulin products. Transfusion. 2015;55(Suppl 2):S117–21. doi:10.1111/trf.13088

Article Snippet: To reduce the levels of isoagglutinins in a chromatographically purified IVIG product (Privigen ® , CSL Behring AG, Bern, Switzerland), an immunoaffinity chromatography (IAC) method was developed; this process was implemented on an industrial scale in 2015.

Techniques: Modification, Chromatography, Fractionation, Diafiltration Assay

Journal: Biodrugs

Article Title: Reduction of Isoagglutinin in Intravenous Immunoglobulin (IVIG) Using Blood Group A- and B-Specific Immunoaffinity Chromatography: Industry-Scale Assessment

doi: 10.1007/s40259-016-0192-3

Figure Lengend Snippet: Anti-A and anti-B titers in intravenous immunoglobulin lots produced with the immunoaffinity chromatography step (Privigen) and intravenous immunoglobulin lots produced using Cohn-like fractionation (Sandoglobulin/Carimune), measured by the European Pharmacopoeia direct method. IAC immunoaffinity chromatography, IgG immunoglobulin G, IVIG intravenous immunoglobulin

Article Snippet: To reduce the levels of isoagglutinins in a chromatographically purified IVIG product (Privigen ® , CSL Behring AG, Bern, Switzerland), an immunoaffinity chromatography (IAC) method was developed; this process was implemented on an industrial scale in 2015.

Techniques: Produced, Chromatography, Fractionation

Journal: Biodrugs

Article Title: Reduction of Isoagglutinin in Intravenous Immunoglobulin (IVIG) Using Blood Group A- and B-Specific Immunoaffinity Chromatography: Industry-Scale Assessment

doi: 10.1007/s40259-016-0192-3

Figure Lengend Snippet: Specific antibody concentrations in intravenous immunoglobulin lots produced with and without the immunoaffinity chromatography step and in intermediates (‘feed’ and ‘flow-through’) of immunoaffinity chromatography

Article Snippet: To reduce the levels of isoagglutinins in a chromatographically purified IVIG product (Privigen ® , CSL Behring AG, Bern, Switzerland), an immunoaffinity chromatography (IAC) method was developed; this process was implemented on an industrial scale in 2015.

Techniques: Produced, Chromatography, Concentration Assay

Journal: Biodrugs

Article Title: Reduction of Isoagglutinin in Intravenous Immunoglobulin (IVIG) Using Blood Group A- and B-Specific Immunoaffinity Chromatography: Industry-Scale Assessment

doi: 10.1007/s40259-016-0192-3

Figure Lengend Snippet: Distribution of immunoglobulin G subclasses in intermediates (‘feed’) before immunoaffinity chromatography and final intravenous immunoglobulin product (‘bulk’)

Article Snippet: To reduce the levels of isoagglutinins in a chromatographically purified IVIG product (Privigen ® , CSL Behring AG, Bern, Switzerland), an immunoaffinity chromatography (IAC) method was developed; this process was implemented on an industrial scale in 2015.

Techniques: Chromatography

Journal: Biodrugs

Article Title: Reduction of Isoagglutinin in Intravenous Immunoglobulin (IVIG) Using Blood Group A- and B-Specific Immunoaffinity Chromatography: Industry-Scale Assessment

doi: 10.1007/s40259-016-0192-3

Figure Lengend Snippet:

Article Snippet: To reduce the levels of isoagglutinins in a chromatographically purified IVIG product (Privigen ® , CSL Behring AG, Bern, Switzerland), an immunoaffinity chromatography (IAC) method was developed; this process was implemented on an industrial scale in 2015.

Techniques: Chromatography, Produced