Production of a human monoclonal IgM directed against human cardiac myosin in a hollow-fiber bioreactor for Membrane Anion Exchange Chromatography one-step purification

Jacobin, Marie-Josée; Santarelli, Xavier; Laroche‐Traineau, Jeanny; Clofent‐Sanchez, Gisèle

doi:10.3233/hab-2004-13303

Cited by 9 publications

(6 citation statements)

References 20 publications

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“…Also knowing the K d of antibody allows judicious selection of antibody for a specific purpose, such as us-ing low affinity antibody in an affinity chromatography procedure for antigen purification [25] or high-affinity antibody in immunoassay techniques [8]. Employment of low affinity antibody in an immunoaffinity column would allow purification and elution of the bound antigen with a mild elution buffer avoiding protein denaturation.…”

Section: Discussionmentioning

confidence: 99%

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Determination of dissociation constants of Anti-ALP monoclonal antibodies by an ELISA – based method

Naseri

Moazzeni

Purfathollah

et al. 2015

HAB

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Section: Discussionmentioning

confidence: 99%

“…Also antibody affinity has been shown to be an important determinant of the efficacy of several laboratory tests, such as hemagglutination, complement fixation, ELISA, precipitation, radioimmunoassay, immunocytochemistry and immunohistochemistry [7][8][9].…”

Section: Introductionmentioning

confidence: 99%

Determination of dissociation constants of Anti-ALP monoclonal antibodies by an ELISA – based method

Naseri

Moazzeni

Purfathollah

et al. 2015

HAB

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“…On the positive side, IgMs are highly charged, and most bind strongly to anion-and cationexchange materials (Jacobin et al 2004), hydroxyapatite (for review, see Gagnon 2009), and weak hydrophobic interaction chromatography media. It is critical to avoid strong hydrophobic surfaces because those have been shown to denature IgM.…”

Section: Purification Of Igmmentioning

confidence: 99%

Antibody Purification and Storage

Fishman¹,

Berg²

2019

Cold Spring Harb Protoc

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Antibodies have become a common and necessary tool in biochemistry, cell biology, and immunology laboratories. There are many different types of antibodies and antibody fragments being used for a myriad of applications. As a result, many different purification protocols have been developed to obtain antibodies of the desired specificity and sensitivity. Here, we introduce the options for small-to largescale antibody purification and isolation of polyclonal and monoclonal antibodies (and fragments generated from these) that target-specific proteins, as well as methods to properly purify antibodies that recognize posttranslational modifications. Optimal conditions for the long-term storage of antibodies are also discussed. ANTIBODY TYPESAntibodies (a.k.a. immunoglobulins [IgGs]) account for 20% of the plasma proteins in humans. Human immunoglobulins consist of several different types of proteins, such as IgD, which functions as an antigen receptor and is found on the surface of B cells; IgE, which binds allergens, subsequently triggering histamine release from mast cells (the "allergic" reaction); IgG, which has four different forms (subclasses IgG1, IgG2, IgG3, and IgG4) (Schur 1987) that provide immunity against foreign pathogens and can also cross the placenta and provide fetal immunity in utero; and IgM, which is unique in that it can exist as a monomer when expressed on the surface of B cells or as a pentamer when it is secreted. IgM represents the initial immune reaction to an infection and, because of its high avidity, can detect and bind even small amounts of antigen. Polyclonal IgG and/or fragments of IgG from various mammalian species such as goats, sheep, or rabbits are used most often for research purposes. Owing to their polyclonal nature, it is not practical in most cases to use polyclonal antibodies for therapeutic applications or for critical applications where large quantities of antibodies will be required for large screens or analyses requiring consistency over a long period of time. Monoclonal antibodies (mAbs) from rodents or rabbits as well as from recombinant sources are the source of most large-scale antibody preparations. Dependent on the source of the antibody, purification and analysis will differ in part based on the final application, requirement for cGMP manufacture, and/or storage conditions required for long-term storage.

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“…(1)(2)(3)(4)(5)(6)(7)(8) Some of these methods are antibody precipitation by, for example, ammonium sulfate (9,10) or polyethylene glycol, (11) gel filtration, (12) ion exchange chromatography, (9,13) and high-performance liquid chromatography (HPLC). (14) Also, general ligand (such as proteins A and G) (15) or specific ligand affinity chromatography (16) can be used.…”

Section: Introductionmentioning

confidence: 99%

Purification of a Murine IgM Monoclonal Antibody

2009

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An IgM monoclonal antibody, S11-23.4, raised against the 47-62 amino acid sequence in bovine prothrombin fragment 1 (F-1, the amino-terminal 156 residues of prothrombin), was purified from tissue culture supernatants and ascites using different purification schemes to determine the best method. There are many different purification schemes for the purification of IgG antibodies, which are generally easier to purify than IgM antibodies. Several different methods and schemes were tried to purify S11-23.4, and it was determined that the best purification schemes are ion exchange chromatography for cell culture IgM antibodies, and a G-100 gel filtration column, in conjunction with precipitation, reduction, and alkylation, for the same IgM antibody in ascites.

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Production of a human monoclonal IgM directed against human cardiac myosin in a hollow-fiber bioreactor for Membrane Anion Exchange Chromatography one-step purification

Cited by 9 publications

References 20 publications

Determination of dissociation constants of Anti-ALP monoclonal antibodies by an ELISA – based method

Determination of dissociation constants of Anti-ALP monoclonal antibodies by an ELISA – based method

Antibody Purification and Storage

Purification of a Murine IgM Monoclonal Antibody

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