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WO1993022438A1 - La proteine l et son procede de preparation par la technologie de l'adn recombine - Google Patents

La proteine l et son procede de preparation par la technologie de l'adn recombine Download PDF

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Publication number
WO1993022438A1
WO1993022438A1 PCT/GB1993/000949 GB9300949W WO9322438A1 WO 1993022438 A1 WO1993022438 A1 WO 1993022438A1 GB 9300949 W GB9300949 W GB 9300949W WO 9322438 A1 WO9322438 A1 WO 9322438A1
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WO
WIPO (PCT)
Prior art keywords
ala
sequence
asn
asp asn
amino acids
Prior art date
Application number
PCT/GB1993/000949
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English (en)
Inventor
Angus Robert Trowern
Antony Atkinson
Jonathan Paul Murphy
Clive James Duggleby
Original Assignee
Microbiological Research Authority
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Microbiological Research Authority filed Critical Microbiological Research Authority
Priority to JP5519105A priority Critical patent/JPH07507925A/ja
Priority to AU42701/93A priority patent/AU674320B2/en
Priority to EP93911929A priority patent/EP0640134A1/fr
Publication of WO1993022438A1 publication Critical patent/WO1993022438A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/33—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Clostridium (G)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide

Definitions

  • This invention relates to novel immunoglobulin binding proteins, processes for their production, recombinant DNA molecules coding therefor and recombinant DNA molecules useful as probes therefor.
  • the present invention relates to the protein designated Protein L in substantially pure and/or intact and/or homogeneous form and to recombinant DNA molecules coding for Protein L.
  • a multitude of Gram-positive bacteria species have been isolated that express surface proteins with affinities for mammalian i munoglobulins through interaction with their heavy chains.
  • the best known of these immunoglobulin binding proteins are type 1 Staphylococcus Protein A and type 2 Streptococcus Protein G which have been shown to interact principally through the C2-C3 interface on the Fc region of human immunoglobulins. In addition, both have also been shown to interact weakly to the Fab region, but again through the immunoglobulin heavy chain.
  • Protein L a novel protein from Peptococcus magrrus , Protein L, has been reported that was found to bind to human, rabbit, porcine, mouse and rat immunoglobulins uniquely through interaction with their light chains. In humans this interaction has been shown to occur exclusively to the kappa chains. Since both kappa and lambda light chains are shared between different classes. Protein L binds strongly to all human classes, in particular to the multi-subunited IgM, and similarly is expected to bind ⁇ to all classes in species that show Protein L light chain binding.
  • Protein L which binds to the Kappa light chain of human immunoglobulins. It has been proposed that Protein L is a virulence factor; non-virulent peptococci and peptostreptococci appear to neither express Protein L nor have the structural gene for it (Kastern et al 1990). Protein L is of particular interest since it has been reported to bind to the Kappa light chain which is present in all classes and sub classes of immunoglobulins. As such it should prove to be a useful diagnostic reagent for use in ELISA and RIA techniques.
  • EP-A-0255 ⁇ 7 describes the purification and attempted characterisation of Protein L by standard protein purification techniques. Subsequently, the authors of EP-A-0255 ⁇ 97 have published a number of scientific papers describing further investigations into the nature and structure of Protein L, but to date, attempts fully to characterize the protein have failed. Thus recently, in a paper entitled "Protein L a Bacterial Immunoglobulin-Binding Protein and Possible Virulence Determinant" by . Kastern et al (Infection and Immunity, May 1990, pp. 1217-1222) there are described unsuccessful attempts to isolate the gene coding for Protein L by determining N-terminal amino acid sequences of tryptic fragments of Protein L and using the derived sequence information to construct probes for isolating the gene.
  • This invention is based on a cDNA sequence comprising a cDNA insert coding for Protein L in its entirety which has now been isolated, thus enabling the above problems to be solved.
  • This cDNA sequence, and the amino acid sequence corresponding to the longest open reading frame thereof, are depicted in Figure 1.
  • the longest open reading frame of the sequence depicted in Figure 1 extends from TTG (103) to AAA(3l83) and the depicted DNA comprises a coding region extending from nucleotide 208 to nucleotide 3183 which codes for immature Protein L.
  • the present invention thus provides a polypeptide designated Protein L and being capable of forming a complex with immunoglobulin Kappa light chains, said polypeptide being characterised by being in substantially homogeneous and/or intact and/or full length form.
  • polypeptides of the invention preferably have at least two and more preferably three of the characterising features of (i) being in substantially homogeneous (ii) being intact and (iii) being in substantially full length form.
  • the full length polypeptides are preferably at least 900 amino acids in length, more preferably are at least 950 amino acids in length and most preferably are at least 975 amino acids in length.
  • the full length polypeptides provided according to the invention include both the mature and immature sequences.
  • the triplet ATG (208) is believed to be the start of the signal sequence which extends for 30 - 35 amino acids.
  • the polypeptides of the invention include both the immature polypeptide extending from Met (208) to Lys (3183) as well as the mature polypeptides which omit 30 to 35 amino acids from the N-terminus.
  • immature protein L or pre-Protein L
  • immature polypeptides of the invention optimally comprise polypeptides which are at least 990 amino acids in length.
  • the immature polypeptides according to the invention preferably have N-terminal sequences corresponding to at least f , more preferably at least 10 and most preferably at least 15 of the amino acids of the N-terminal sequence Met Lys lie Asn Lys Lys Leu Leu Met Ala Ala Leu Ala Gly Ala He Val Val Gly Gly
  • N-terminal sequences are:
  • the invention further includes polypeptides as defined above, but omitting a signal sequence.
  • the signal sequence is between 20 and 35 amino acids in length, more specifically, between 23 and 27 amino acids in length.
  • the full length mature polypeptides of the invention preferably commence with one of the following N-terminal sequences: Gly Ala Asn Ala Tyr Ala Ala Glu Glu Asp Asn Thr Asp Asn Asn ...
  • the invention also includes variants of the polypeptides defined above, all variants being capable of forming a complex with immunoglobulin Kappa light chains.
  • the variant polypeptides of the invention have at least 7 sequence homology, preferably at least 0 sequence homology with the amino acid sequence depicted in Figure 1. Most preferably they have at least 95% sequence homology, preferably at least 98% sequence homology with the amino acid sequence depicted in Figure 1.
  • the full length polypeptide depicted in Figure 1 has the C-terminal sequence Leu Ala Ala Ala Ala Leu Ser Thr Ala Ala Gly Ala Tyr Val Ser Leu Lys Lys Arg Lys.
  • Polypeptides according to the invention preferably have C-terminal sequences corresponding to at least 7. more preferably at least 10 and most preferably at least 15 of the amino acids of the C-terminal sequence Leu Ala Ala Ala Ala Leu Ser Thr Ala Ala Gly Ala Tyr Val Ser Leu Lys Lys Arg Lys.
  • the invention further provides, in a second aspect, recombinant DNA molecules having an insert coding for polypeptides as defined above.
  • recombinant DNA molecules which code for the immature polypeptides of the invention are selected from (a) the DNA coding sequence depicted in Figure 1 commencing with ATG in location 208 and extending to AAA in location 3183. and (b) degenerate DNA sequence coding for the same amino acid sequence.
  • the recombinant DNA molecules which code for the mature polypeptides of the invention are selected from (c) the DNA coding sequence depicted in Figure 1 commencing with a codon in the region between locations 211 and 313 and extending to AAA In location 3183, and (d) degenerate DNA sequence coding for the same amino acid sequence.
  • the present invention provides a process for producing an immunoglobulin binding protein which comprises culturing a transformed host which has been transformed with an expression vector containing a DNA coding sequence as defined above.
  • the transformed host may be eukaryotic or prokaryotic, e.g. a bacteria or yeast or an animal cell, a cultured mammalian cell or an Insect cell.
  • the invention further provides, in a fourth aspect, recombinant DNA molecules comprising a contiguous sequence of at least 12, preferably 15, and most preferably 17 bases depicted In the DNA sequence of Figure l and being useful as probes for isolating other DNA sequences coding for immunoglobulin binding protein.
  • the DNA sequence depicted in Figure 1 is used to construct probes useful in probing gene banks for DNA sequences coding for polypeptides of related sequence to Protein L.
  • the present invention further provides, in a fifth aspect, a process for producing an immunoglobulin binding protein which comprises culturing an expression vector containing a DNA coding sequence isolated by probing- a gene bank with a probe as defined above.
  • the specific binding properies of Protein L including its ability to bind immunoglobulin Kappa light chains, is believed to be attributable to the presence of sequences which have a recognisably repeated character within the amino acid sequence of the molecule.
  • the term "recognisably repeated character" as used herein is meant that the amino acid sequence comprises at least two sequences, each of from 20 to 45 amino acids in length, which have an at least 75%. preferably at least 90% and most preferably at least 95% homology with one another.
  • the polypeptide sequence depicted in Figure 1 includes ten sets of repeated sequences at least two of which are considered to be responsible for immunoglobulin Kappa light chain binding.
  • the ability to bind Kappa light chains is considered to be associated with one or more of the repeated sequences A, B, C and D (sequences (1) - (4) above). —
  • synthetic immunoglobulin binding molecules comprising a plurality of recognisably repeated binding domains selected from the sequences which are labelled at their N-terminal ends in Figure 1 as Al, A2 and A3; Bl, and B2; Cl, C2, C3. and C4; and Dl, D2, D3 and D4.
  • the synthetic immunoglobulin binding molecules preferably comprise from 2 to 1 of said domains.
  • the selected domain or domains may be identical to the sequences which are labelled at their N-terminal ends in Figure 1 as Al, A2 and A3; Bl, and B2; Cl, C2, C3, and C4; Dl, D2, D3 and D4, or they may vary from said sequences, provided that they have an at least 75% " . preferably at least 90% and most preferably at least 95% homology therewith.
  • the sequences labelled at their N-terminal ends as El. E2 and E3; and FI.
  • F2, F3 and F4 are believed to be responsible for albumin binding and the synthetic binding molecules provided according to the Invention may include sequences selected from sequences El, E2 and E3; and FI, F2, F3 and F4 or related squences which vary from said sequences, provided that they have an at least 75%. preferably at least 90% and most preferably at least 95% homology therewith.
  • a collection of 56 different clinical isolates of Gram-positive anaerobic cocci were obtained from the Luton Public Health Laboratory Anaerobe Reference Unit, Bath Public Health Laboratory and Salisbury Public Health Laboratory.
  • Type strains of peptococci and peptostreptococci were from the National Collection of Type Cultures, Central Public Health Laboratory, Colindale, London.
  • Proteins from the cell extracts were separated by electrophoresis on 7-5% polyacrylamide SDS gels, and electrophoretically transferred to nitrocellulose membranes (Hybond C; Amersham) by Western blotting (Harlow & Lane, 1988) . After overnight transfer, the membranes were washed several times in phosphate-buffered saline containing 0.02% Tween 20 (PBS-T) and then the protein binding sites were blocked by incubation in 1% gelatine (Sigma) in PBS-T for one hour.
  • PBS-T phosphate-buffered saline containing 0.02% Tween 20
  • Immunoglobulin-binding proteins were then detected by incubation with either alkaline phosphatase-coupled human IgG or phosphatase-coupled light chain (prepared from human IgG by reduction, alkylation and separation by FPLC - Harlow and Lane, 1988) . Immunoglobulin-binding proteins were detected using nitro-blue tetrazolium and X-phos (Harlow & Lane, 1988) .
  • a 4-litre culture of strain 1018 was grown up, harvested and lysed as described above.
  • the cell extract was heat-treated (80°C, 10 min) prior to the centrifugation step.
  • the supernatant was made to 250 mM NaCl and applied to a 5ml column of IgG-Sepharose (15 mm diameter).
  • the column was washed with 50 M Hepes-NaOH, pH 8.0, and immunoglobulin-binding proteins were eluted with lOOmM glycine-HCl, pH 2.0.
  • the protein-containing eluate was neutralised with Tris-HCl to pH 7-5 prior to analysis by Western blotting.
  • the sizes of the immunoglobulin-binding proteins in the P. magnus isolates corresponds with the reported size of Protein L.
  • the clinical isolates that had immunoglobulin-binding proteins were from wound isolates or infected surgical wounds.
  • the immunoglobulin-binding proteins were detected using an alkaline phosphatase-labelled light chain preparation from human IgG. 1. Conclusions.
  • Radiochemicals were from Amersham International. X-Omat S X-ray film was from Kodak. Deoxy- and dideoxy-nucleoside triphosphates, DNA ligase, restriction endonucleases and other DNA-modifying enzymes were from Boehringer. Agarose, acrylamide, bisacrylamide and phenol were from Bethesda Research Laboratories. Chromatography media were from Pharmacla-LKB (Uppsala, Sweden) . Human Immunoglobulins and serum albumin were from Sigma. All other reagents were from Sigma or BDH. NitroceHulose was purchased from Anderman and Co. , Singer-upon- Thames, Surrey, U.K.
  • E.coli TGI was cultured in 2xYT broth (2% (w/v) try ⁇ tone/1% (w/v) yeast extract/1% (w/v) NaCl) overnight at 37 ° C. Media were solidified with 2% (w/v) Bacto-agar (Difco) .
  • HT-agar for M13 overlays contained 1% (w/v) tryptone, 0.8% (w/v) NaCl and 0.8% (w/v) Bacto-agar (Difco) . Ampicillin at a concentration of 50 ⁇ g/ml was used where necessary for the selection and growth of transformants.
  • ⁇ -galactosidase was detected by addition of 5-bromo-4-chlorindolyl- ⁇ - D-galactoside to a final concentration of 600 ⁇ g/ml and, where necessary, isopropyl- ⁇ -D-thiogalactopyranoside to a final concentration of 200 ⁇ g/ml.
  • Plasmids and phage RF DNA were purified from E. co l i by Brij lysis and CsCl/ethidium bromide density-gradient centrifugation. Peptococcus chromosomal DNA was isolated as described elsewhere.
  • DNA-modifying enzymes were used in the buffer and under the conditions recommended by the supplier (Boehringer) . Transformation of E. co li was essentially as described previously. Electrophoresis of DNA fragments was performed on vertical 1% (w/v)-agarose slab gels in Tris-acetate buffer (40 mM-Tris/20 mM-sodium acetate/2 mM-EDTA, adjusted to pH 7-9 with acetic acid). DNA fragment sizes were estimated by comparison with fragments of lambda phage DNA previously digested with the restriction endonuclease Hind III. DNA fragments were purified by electroelution essentially as described previously.
  • Nucleotide sequences were determined by the chain- ermination procedure on M13 templates using a shotgunning protocol to generate random templates. Multiple overlapping sequences were compiled using the programmes supplied by DNASTAR Inc (Maidison, U.S.A.). These same programmes were used to analyse the sequenced gene and its translated protein. Oligonucleotide primers were synthesised by using the Applied biosysterns 380B DNA synthesiser.
  • a cell suspension was transferred to a MSE sonication tube and subjected to ultra sonication (3 ⁇ 30 sec bursts at 18MH with 30 sec intervals, at 4 C using an MSE Soniprep 150 Sonicator) .
  • a sonication procedure was used to disrupt bacterial cells from small scale purification of PPL by affinity chromatography on IgG-sepharose FF. Cultures of 300ml were grown overnight then centrifuged (15000 g for 10 min at 4 ° C) and resuspended in 3 ml of 100 mM Tris-HCl, pH 7-5. 250 mM NaCl. The suspension was sonicated, centrifuged (30000 g 10 min at 4°C) and the supernatant fluid passed through a 1ml column (1.6 cm x 0.90 cm i.d.) of IgG-Sepharose FF equilibrated and washed with 5ml of 100 mM tris-HCl, pH 7-5. 250 mM NaCl. The protein was eluted with 100 mM glycine-HCl, pH 2.0, and the pH raised to 7-5 using 1M tris, pH 8.0.
  • Proteins were applied to nitrocellulose membranes by electrophoretic transfer from SDS-polyacrylamide gels and probed with
  • N-terminal amino acid sequences were determined on an Applied Biosystems 77A pulsed " liquid protein sequencer by automated Edman - phenylthiohydantoin degradation. PPL samples were dialysed against 0 mM-NaCl, and about 500 pmol was applied to the gas-phase sequencer. The equipment was operated essentially according to the manu acturer's instructions. Repetitive Edman degradations provided sequential removal of amino acids from the peptide, which were identified by using reversed-phase HPLC.
  • ⁇ PPL9 was found to contain the smallest insert of 6.2Kb, and through sub-cloning the ppl gene was narrowed down to a 4.2Kb Pstl fragment. This Pstl fragment was subsequently excised, and sonicated by shotgunning.
  • Protein L can form the basis of numerous systems where Kappa light chain binding is desired.
  • Protein L can be used as a reagent for immobilising antibodies, e.g. on columns, in diagnostic tests and in assays. Additional uses are as pharmaceuticals and as reagents for preparing pharmaceuticals.
  • Synthetic immunoglobulin binding molecules according to the invention can also form the basis of numerous systems where Kappa light chain binding is desired, e.g. test kits, biochemical reagents, protocols.
  • these synthetic molecules may omit sequences selected from sequences El, E2, E3, FI, F2 and F3 so as to be substantially free of albumin binding ability.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
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Abstract

L'invention se rapporte à la protéine L se présentant sous une forme allongée pratiquement homogène, et/ou intacte, et/ou de longueur totale, à son procédé de fabrication, à l'ADN recombiné la codant et aux molécules synthétiques dérivées de celle-ci.
PCT/GB1993/000949 1992-05-07 1993-05-07 La proteine l et son procede de preparation par la technologie de l'adn recombine WO1993022438A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
JP5519105A JPH07507925A (ja) 1992-05-07 1993-05-07 プロテインｌおよび組換えｄｎａ法によるその製法
AU42701/93A AU674320B2 (en) 1992-05-07 1993-05-07 Protein l and process for its preparation by recombinant dna technology
EP93911929A EP0640134A1 (fr) 1992-05-07 1993-05-07 La proteine l et son procede de preparation par la technologie de l'adn recombine

Applications Claiming Priority (2)

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GB929209804A GB9209804D0 (en) 1992-05-07 1992-05-07 Novel immunoglobulin binding proteins process for their production and recombinant dna molecules coding therefor
GB9209804.5 1992-05-07

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JP (1) JPH07507925A (fr)
AU (1) AU674320B2 (fr)
CA (1) CA2135207A1 (fr)
GB (1) GB9209804D0 (fr)
WO (1) WO1993022438A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997035887A1 (fr) * 1996-03-26 1997-10-02 Amrad Operations Pty. Ltd. Precurseurs d'anticorps catalytiques
WO1998034956A1 (fr) * 1997-02-07 1998-08-13 Commissariat A L'energie Atomique Complexe non-covalent comprenant au moins un anticorps et un element de liaison aux immunoglobulines associe a une substance active, son procede de preparation et ses applications
WO1999015563A1 (fr) * 1997-09-19 1999-04-01 Amrad Operations Pty. Ltd. Anticorps catalytiques et leur mode de production
EP3375873A4 (fr) * 2015-10-22 2019-05-22 Protenova Co., Ltd. Polypeptide se liant aux immunoblobulines

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ATE198768T1 (de) * 1992-05-07 2001-02-15 Affitech As Von l protein abgeleitete immunoglobulin-bindende proteine und deren anwendungen

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0255497A2 (fr) * 1986-07-22 1988-02-03 Hightech Receptor Ab Protéine L et ses sous-fragments, à activité de liaison d'immunoglobuline, procédé de préparation, trousse de réactif, composition pharmaceutique et souche de Peptococcus magnus

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ATE198768T1 (de) * 1992-05-07 2001-02-15 Affitech As Von l protein abgeleitete immunoglobulin-bindende proteine und deren anwendungen

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0255497A2 (fr) * 1986-07-22 1988-02-03 Hightech Receptor Ab Protéine L et ses sous-fragments, à activité de liaison d'immunoglobuline, procédé de préparation, trousse de réactif, composition pharmaceutique et souche de Peptococcus magnus

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
INFECTION AND IMMUNITY vol. 58, no. 5, May 1990, WASHINGTON US pages 1217 - 1222 Kastern W;Holst E;Nielsen E;Sjobring U;Bjorck L; 'Protein L, a bacterial immunoglobulin-binding protein and possible virulence determinant.' *
JOURNAL OF BIOLOGICAL CHEMISTRY vol. 264, no. 33, 25 November 1989, BALTIMORE, MD US pages 19740 - 19746 Akerstrom B;Bjorck L; 'Protein L: an immunoglobulin light chain-binding bacterial protein. Characterization of binding and physicochemical properties.' *
JOURNAL OF BIOLOGICAL CHEMISTRY vol. 267, no. 18, 25 June 1992, BALTIMORE, MD US pages 12820 - 12825 Kastern W;Sjobring U;Bjorck L; 'Structure of peptostreptococcal protein L and identification of a repeated immunoglobulin light chain-binding domain.' *
JOURNAL OF BIOLOGICAL CHEMISTRY vol. 267, no. 4, 5 February 1992, BALTIMORE, MD US pages 2234 - 2239 Nilson BH;Solomon A;Bjorck L;Akerstrom B; 'Protein L from Peptostreptococcus magnus binds to the kappa light chain variable domain.' *

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997035887A1 (fr) * 1996-03-26 1997-10-02 Amrad Operations Pty. Ltd. Precurseurs d'anticorps catalytiques
GB2326643A (en) * 1996-03-26 1998-12-30 Amrad Operations Pty Ltd Precursors of catalytic antibodies
GB2326643B (en) * 1996-03-26 2000-09-27 Amrad Operations Pty Ltd Precursors of catalytic antibodies
WO1998034956A1 (fr) * 1997-02-07 1998-08-13 Commissariat A L'energie Atomique Complexe non-covalent comprenant au moins un anticorps et un element de liaison aux immunoglobulines associe a une substance active, son procede de preparation et ses applications
FR2759296A1 (fr) * 1997-02-07 1998-08-14 Commissariat Energie Atomique Complexe non-covalent comprenant au moins un anticorps et un element de liaison aux immunoglobulines associe a une substance active, son procede de preparation et ses applications
WO1999015563A1 (fr) * 1997-09-19 1999-04-01 Amrad Operations Pty. Ltd. Anticorps catalytiques et leur mode de production
GB2345694A (en) * 1997-09-19 2000-07-19 Amrad Operations Pty Ltd Catalytic antibodies and a method of producing same
GB2345694B (en) * 1997-09-19 2002-07-17 Amrad Operations Pty Ltd Catalytic antibodies and a method of producing same
EP3375873A4 (fr) * 2015-10-22 2019-05-22 Protenova Co., Ltd. Polypeptide se liant aux immunoblobulines
US11208441B2 (en) 2015-10-22 2021-12-28 Protenova Co., Ltd. Immunoglobulin-binding polypeptide

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GB9209804D0 (en) 1992-06-24
AU4270193A (en) 1993-11-29
JPH07507925A (ja) 1995-09-07
CA2135207A1 (fr) 1993-11-11
EP0640134A1 (fr) 1995-03-01
AU674320B2 (en) 1996-12-19

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