公开(公告)号：WO2020124252A1

公开(公告)日：2020-06-25

申请号：PCT/CA2019/051870

申请日：2019-12-19

申请人： RAPID NOVOR INC.

发明人： LE BIHAN, Thierry ,  MA, Bin ,  MCDONALD, Zac ,  LIU, Qixin ,  TAYLOR, Paul

IPC分类号： G01N33/483 ,  C12Q1/37 ,  G01N1/28 ,  C07K1/107 ,  C07K1/10

摘要： A method for increasing peptide fragmentation by labelling the peptide at the C-terminal end with a guanidinium group or other basic functional group and distinguishing isobaric amino acids and amino acid combinations of asparagine and glycine-glycine; glutamine and glycine-alanine; and/or glutamine and alanine-glycine, during polypeptide sequencing. The method involves: obtaining a peptide of interest and/or digesting a polypeptide of interest with a protease, such as pepsin, chymotrypsin or trypsin, or by chemical cleavage to produce shorter peptides; reacting the obtained and/or generated peptides with a coupling reagent to derivatize the free C-terminal carboxylic acid function of the peptides, thus adding a basic functional group rendering C-terminal peptide fragment ions detectable by mass spectrometry; selecting a charge state of 2+ or more, and fragmenting the derivatized peptides in a mass spectrometer under conditions effective to generate at least w ions; and detecting the w ions by mass spectrometry, and identifying derivatized peptides which incorporate the additional mass of the basic functional group.

公开(公告)号：WO2023010219A1

公开(公告)日：2023-02-09

申请号：PCT/CA2022/051194

申请日：2022-08-05

申请人： RAPID NOVOR, INC.

发明人： LE BIHAN, Thierry ,  NUNEZ DE VILLAVICENCIO DIAZ, Teresa ,  LIYASOVA, Mariya ,  YAO, Chenyu ,  LIU, Qixin ,  REITZEL, Chelsea ,  DUAN, Jin ,  TAYLOR, Paul ,  MA, Bin

IPC分类号： C12Q1/37 ,  C07K1/14 ,  C07K1/18 ,  C07K16/00 ,  G01N30/02 ,  G01N30/60 ,  G01N30/72 ,  G01N33/53 ,  G01N33/543

摘要： The present application relates to methods for determining antibody heavy and light chain pairing in an antibody and/or antibody fragment mixture. The methods include a step of submitting the mixture to a separation step under non-reducing condition to obtain isolated antibodies or antibody fragments, or of contacting the mixture with a protein cross-linking agent. The isolated or cross-linked antibodies and/or antibody fragments are then digested using one or more proteases to generate antibody or antibody fragment peptides, which are then analyzed for example by mass spectrometry (MS) to determining heavy chain and light chain pairing of the antibody or antibody fragment in the mixture.

公开(公告)号：WO2021248244A1

公开(公告)日：2021-12-16

申请号：PCT/CA2021/050791

申请日：2021-06-10

申请人： RAPID NOVOR, INC.

发明人： LE BIHAN, Thierry ,  MA, Bin ,  TAYLOR, Paul ,  YAO, Chenyu ,  LIU, Qixin ,  REITZEL, Chelsea ,  GOROSPE, Kayle Kathleen Marie ,  LIYASOVA, Mariya

IPC分类号： C07K16/42 ,  C07K1/107 ,  C07K1/22 ,  C07K16/00 ,  C07K7/06 ,  C07K7/08 ,  C12P21/06 ,  C07K16/06 ,  C07K16/065 ,  C07K2317/34 ,  C07K2317/565

摘要： Methods and reagents to obtaining a sample enriched in peptides comprising the third complementarity-determining region of the heavy chain (CDRH3) of immunoglobulins, such as IgGs, are described. These methods are based on the use of targeted protease digestion of immunoglobulins and affinity purification of CDRH3 peptides using specific antibodies. Such methods and reagents are useful for analyzing the immunoglobulin repertoire.