Montanide isa 51 adjuvant

Manufactured by Seppic

Sourced in France

Montanide ISA-51 is a water-in-oil (W/O) emulsion adjuvant. Its core function is to enhance and modulate the immune response generated by vaccines.

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Lab products found in correlation

Imiquimod, by 3M (1 mentions)

6 protocols using montanide isa 51 adjuvant

Immunization of Mice and Rabbits with MERS-CoV RBD

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Mice were immunized with MERS-CoV RBD fragments following a previously described protocol [18 (link),29 (link)]. Briefly, mice were prime-vaccinated (subcutaneously, s.c.) with 10 μg/mouse of recombinant MERS-CoV RBD fragments in the presence of Montanide ISA 51 adjuvant (SEPPIC, Fairfield, NJ), and boosted twice with the same immunogen and adjuvant at 3-week (21 days) intervals. Sera from 10 days post-last vaccination (52 days) were heat-inactivated at 56°C for 30 minutes, and detected for MERS-CoV S-specific antibody response and neutralizing antibodies. Sera from 0, 10, 31, and 136 days post-initial immunization of S377-588-Fc were also tested for MERS-CoV S-specific antibody response.
The rabbit immunization (Covance Research Products Inc., Denver, PA) followed a protocol similar to that of mice with some modifications. Briefly, rabbits were prime-vaccinated with MERS-CoV S377-588-Fc fragment (250 μg/rabbit) plus Freund's complete adjuvant and then boosted twice at 3-week intervals with the same immunogen (125 μg/rabbit) plus Freund's incomplete adjuvant. Sera were collected and tested as above.

Ma C., Wang L., Tao X., Zhang N., Yang Y., Tseng C.T., Li F., Zhou Y., Jiang S, & Du L. (2014). Searching for an ideal vaccine candidate among different MERS coronavirus receptor-binding fragments—The importance of immunofocusing in subunit vaccine design. Vaccine, 32(46), 6170-6176.

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Immunization protocol for MERS-CoV RBD fragments

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Mice were immunized with MERS-CoV RBD fragments following a previously described protocol [18] (link), [29] (link). Briefly, mice were prime-vaccinated (subcutaneously, s.c.) with 10 μg/mouse of recombinant MERS-CoV RBD fragments in the presence of Montanide ISA 51 adjuvant (SEPPIC, Fairfield, NJ), and boosted twice with the same immunogen and adjuvant at 3-week (21 days) intervals. Sera from 10 days post-last vaccination (52 days) were heat-inactivated at 56 °C for 30 min, and detected for MERS-CoV S-specific antibody response and neutralizing antibodies. Sera from 0, 10, 31, and 136 days post-initial immunization of S377-588-Fc were also tested for MERS-CoV S-specific antibody response.
The rabbit immunization (Covance Research Products Inc., Denver, PA) followed a protocol similar to that of mice with some modifications. Briefly, rabbits were prime-vaccinated with MERS-CoV S377-588-Fc fragment (250 μg/rabbit) plus Freund's complete adjuvant and then boosted twice at 3-week intervals with the same immunogen (125 μg/rabbit) plus Freund's incomplete adjuvant. Sera were collected and tested as above.

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Peptide-based Influenza Vaccine Formulation

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FLU-v is a lyophilized vaccine composed of four short peptides; FLU-5 (32aa), FLU-7 (21aa), FLU-8N (20aa), and FLU-10 (24aa)^{19 (link),20 (link)}. These peptides originate from conserved regions in internal proteins M1, NPA, NPB, and M2 respectively. The peptides are manufactured in the solid phase by fluorenylmethoxycarbonyl (Fmoc) chemistry, reconstituted, filtered, and lyophilized. It was administered by subcutaneous injection as a water-in-oil emulsion (50:50) with water for injection and Montanide ISA-51 adjuvant (Seppic, France) with 500 μg FLU-v in a final volume of 0.5 ml. The placebo was prepared by emulsifying 0.25 ml of water for injection and 0.25 ml of Montanide ISA-51. Montanide ISA-51 is a ready to use adjuvant composed of light mineral oil and a surfactant (mannide monooleate from vegetable oil) designed to form water-in-oil emulsions in order to enhance the immune response against targeted antigens.

Pleguezuelos O., James E., Fernandez A., Lopes V., Rosas L.A., Cervantes-Medina A., Cleath J., Edwards K., Neitzey D., Gu W., Hunsberger S., Taubenberger J.K., Stoloff G, & Memoli M.J. (2020). Efficacy of FLU-v, a broad-spectrum influenza vaccine, in a randomized phase IIb human influenza challenge study. NPJ Vaccines, 5, 22.

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Adjuvant-assisted Immunotherapy Protocol

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GM-CSF (Leukine) was manufactured by Berlex (Seattle, Washington, USA) as a clinical grade reagent, and was purchased from the UVA hospital pharmacy. Montanide ISA-51 adjuvant was purchased from Seppic (Paris, France) in sealed glass vials (3 mL/vial). Montanide ISA-51 had been prepared with oleic acid from a bovine source (‘animal’ formulation) for 23 participants and from a vegetable source for 5 participants.37 (link) DMSO (Cryoserv) was purchased from the manufacturer (Edwards Lifesciences Research Medical.; Irvine, California, USA). Imiquimod (Aldara; 5% cream, 0.25 g/packet) was manufactured by 3M (St. Paul, Minnesota, USA) as a clinical grade reagent and was purchased from the UVA hospital pharmacy.

Meneveau M.O., Petroni G.R., Salerno E.P., Lynch K.T., Smolkin M., Woodson E., Chianese-Bullock K.A., Olson W.C., Deacon D., Patterson J.W., Grosh W.W, & Slingluff C.L. (2021). Immunogenicity in humans of a transdermal multipeptide melanoma vaccine administered with or without a TLR7 agonist. Journal for Immunotherapy of Cancer, 9(5), e002214.

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Experimental Melanoma Vaccine Trial

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Thirty-seven evaluable participants with American Joint Committee on Cancer stage IIIB, IIIC or IV melanoma with (n=17) or without (n=20) measurable disease were vaccinated with an experimental melanoma vaccine comprised of 6 peptides 14-23 amino acids in length (Table 1). The patients were required to express HLA-DR1, DR4, DR11, DR13, or DR15. The peptides were administered with 110 mcg GM-CSF (Berlex, Seattle, WA) in a stable emulsion with 1 ml Montanide ISA-51 adjuvant (IFA, Seppic, Inc., Paris, France/ Fairfield, NJ) at weeks 0, 1, 2, 4, 5, and 6. Following the third vaccination, a sentinel immunized node (SIN) draining the replicate vaccine site was removed surgically. Additional enrollment and study design details have been reported previously [7 (link)]. Patients’ blood and tissue samples were studied following informed consent, and with Institutional Review Board (HIC#10464) and FDA approval (BB-IND #1825).

Hu Y., Petroni G.R., Olson W.C., Czarkowski A., Smolkin M.E., Grosh W.W., Chianese-Bullock K.A, & Slingluff CL J.r. (2014). Immunologic Hierarchy, Class II MHC Promiscuity, and Epitope Spreading of a Melanoma Helper Peptide Vaccine. Cancer immunology, immunotherapy : CII, 63(8), 779-786.

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Immunization with Recombinant Antigen

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C57BL/6 mice (n = 4) received 3 subcutaneous injections with 20 μg nFhGST mixed with an equal amount of water-in-oil emulsion Montanide ISA51 adjuvant (Seppic Inc, Fairfield, NJ). The injections were applied two weeks apart. Animals were bled out one week after the last injection and the sera from each individual animal were pooled. Anti-serum had antibody titers of ~1: 25,600 when titrated by ELISA against nFhGST.

Aguayo V., Valdés Fernandez B.N., Rodríguez-Valentín M., Ruiz-Jiménez C., Ramos-Benítez M.J., Méndez L.B, & Espino A.M. (2019). Fasciola hepatica GST downregulates NF-κB pathway effectors and inflammatory cytokines while promoting survival in a mouse septic shock model. Scientific Reports, 9, 2275.

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