The present invention concerns a method of storing a separation matrix comprising multimers of immunoglobulin-binding alkali-stabilized Protein A domains covalently coupled to a porous support. The method comprises the steps of: a) providing a storage liquid comprising at least 50% by volume of an aqueous alkali metal hydroxide solution; b) permeating the separation matrix with the storage liquid; and c) storing the storage liquid-permeated separation matrix for a storage time of at least days. The alkali-stabilized Protein A domains comprise mutants of a parental Fc-binding domain of Staphylococcus Protein A (SpA), as defined by, or having at least 80% such as at least 90%, 95% or 98% identity to, SEQ ID NO 51 or SEQ ID NO 52, wherein the amino acid residues at positions 13 and 44 of SEQ ID NO 51 or 52 are asparagines and wherein at least the asparagine residue at position 3 of SEQ ID NO 51 or 52 has been mutated to an amino acid selected from the group consisting of glutamic acid, lysine, tyrosine, threonine, phenylalanine, leucine, isoleucine, tryptophan, methionine, valine, alanine, histidine and arginine.

The present invention relates to a chromatography ligand, which comprises Domain C from Staphylococcus protein A (SpA), or a functional fragment or variant thereof. The chromatography ligand presents an advantageous capability of withstanding harsh cleaning in place (CIF) conditions, and is capable of binding Fab fragments of antibodies. The ligand may be provided with a terminal coupling group, such as arginine or cysteine, to facilitate its coupling to an insoluble carrier such as beads or a membrane. The invention also relates process of using the ligand in isolation of antibodies, and to a purification protocol which may include washing steps and/or regeneration with alkali.

The present invention relates to a chromatography ligand, which comprises Domain C from Staphylococcus protein A (SpA), or a functional fragment or variant thereof. The chromatography ligand presents an advantageous capability of withstanding harsh cleaning in place (CIF) conditions, and is capable of binding Fab fragments of antibodies. The ligand may be provided with a terminal coupling group, such as arginine or cysteine, to facilitate its coupling to an insoluble carrier such as beads or a membrane. The invention also relates process of using the ligand in isolation of antibodies, and to a purification protocol which may include washing steps and/or regeneration with alkali.

An Fc-binding polypeptide of improved alkali stability, comprising a mutant of an Fc-binding domain of Staphylococcus Protein A (SpA), as defined by SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO:3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO 26 or SEQ ID NO 27, wherein at least the alanine residue at the position corresponding to position 42 in SEQ ID NO:4-7 has been mutated to arginine and/or wherein at least the aspartic acid residue at the position corresponding to position 37 in SEQ ID NO:4-7 has been mutated to glutamic acid.

An Fc-binding polypeptide of improved alkali stability, comprising a mutant of an Fc-binding domain of Staphylococcus Protein A (SpA), as defined by SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO:3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO 26 or SEQ ID NO 27, wherein at least the alanine residue at the position corresponding to position 42 in SEQ ID NO:4-7 has been mutated to arginine and/or wherein at least the aspartic acid residue at the position corresponding to position 37 in SEQ ID NO:4-7 has been mutated to glutamic acid.

The present invention relates to the field of protein purification and relates in particular to novel proteins that bind specifically to acid alpha glucosidase (GAA). The invention further relates to fusion proteins comprising novel proteins that bind specifically to GAA. In addition, the invention relates to affinity matrices comprising the GAA binding proteins of the invention. The invention also relates to a use of these GAA binding proteins or affinity matrices for affinity purification of GAA and to methods of affinity purification of GAA using the GAA binding proteins of the invention. Further uses relate to analytical methods for the determination of GAA in liquids.

The present invention relates to the field of protein purification and relates in particular to novel proteins that bind specifically to acid alpha glucosidase (GAA). The invention further relates to fusion proteins comprising novel proteins that bind specifically to GAA. In addition, the invention relates to affinity matrices comprising the GAA binding proteins of the invention. The invention also relates to a use of these GAA binding proteins or affinity matrices for affinity purification of GAA and to methods of affinity purification of GAA using the GAA binding proteins of the invention. Further uses relate to analytical methods for the determination of GAA in liquids.

The invention relates to methods for modulating the neurological and immune function through targeting of acetylcholine (ACh) and CLIP. The result is wide range of new therapeutic regimens for treating, inhibiting the development of, or otherwise dealing with, a multitude of illnesses and conditions, including psychiatric or neurological disease, autoimmune disease, transplant and cell graft rejection, chronic wounds, non-neuronal immune disorders, Alzheimer's, Parkinson's Disease, Lewy Body dementia, pediatric acute neuropsychiatric syndromes (PANS), hypertension, late stage Ebola, Hantavirus, or coronavirus-induced hyperinflammatory conditions, including infections that cause a pathological “cytokine storm”, other post-infectious syndromes that result in cytokine storms in some people, and cancer, as well as novel methods of diagnosis and of introducing a treatment regimen into a subject and improving cognition and addiction cessation methods. In some embodiments the addiction is an addiction to smoking, alcohol and/or drugs.

The invention relates to methods for modulating the neurological and immune function through targeting of acetylcholine (ACh) and CLIP. The result is wide range of new therapeutic regimens for treating, inhibiting the development of, or otherwise dealing with, a multitude of illnesses and conditions, including psychiatric or neurological disease, autoimmune disease, transplant and cell graft rejection, chronic wounds, non-neuronal immune disorders, Alzheimer's, Parkinson's Disease, Lewy Body dementia, pediatric acute neuropsychiatric syndromes (PANS), hypertension, late stage Ebola, Hantavirus, or coronavirus-induced hyperinflammatory conditions, including infections that cause a pathological “cytokine storm”, other post-infectious syndromes that result in cytokine storms in some people, and cancer, as well as novel methods of diagnosis and of introducing a treatment regimen into a subject and improving cognition and addiction cessation methods. In some embodiments the addiction is an addiction to smoking, alcohol and/or drugs.

The disclosure provides modified biotin-binding proteins which can be expressed in soluble form in high yield in bacteria. Also provided are fusion proteins comprising the modified biotin-binding protein and an antigen. The disclosure further provides non-hemolytic variants of alpha-hemolysin from S. aureus and fusion protein comprising non-hemolytic variant of alpha-hemolysin and a biotin-binding domains. Immunogenic compositions comprising the proteins are also disclosed and use of such immunogenic compositions for inducing an immune response or for vaccinating a subject are also disclosed.