Required information for biologics documents

Firms requesting a USAN for a protein, monoclonal antibody, oligonucleotide based substance, cell therapy, or gene therapy are required to include an MS Word document with their application to further define the substance and its structural information. The information required in this document depends on the type of substance. 

USAN Requirements are aligned with those of the World Health Organization’s INN Expert Committee. Find more details on their guidelines.

All proteins

• Complete mature amino acid sequence as editable text, using single-letter codes for each amino acid with spaces between groups of ten characters, five groups per line and with a number indicating the position of the last amino acid at the end of each line.
• Complete precursor nucleotide sequence with spaces between codons and translation (including the stop codon) and with numbers per line, as editable text.
• If applicable, state and explain the purpose of amino acid differences with the native sequence.
• Positions of all disulfide bridges (specify if they are determined or predicted).
• Post‐translational modifications (specify if they are determined or predicted).
• Expression system (the cell type, specific strain, and the clone name used for the expression).
• If available, the three-dimensional structure in Protein Data Bank format or the Protein Data Bank accession code.
• Glycosylated proteins. The glycosylation profile (the types of sugar, the location of glycosylation site(s), etc.); specify if they are determined or predicted; if the cell line in which the protein/peptide is produced is engineered, detailed information if the glycosylation pattern is affected.
• Conjugates. The mean numbers of molecules of the conjugated part, and if known, positions where the conjugate is attached.

Monoclonal antibodies, antibody fragments, multi-specific antibodies, antibody drug conjugates

• All information required for all proteins and peptides.
• CDR-IMGT and sequence analysis of the variable regions showing percentage of human content.
• CDR-Kabat (sequence and residue range).
• IG class and subclass, IG format and chain types.
• Species or taxonomy related structure (chimeric, humanized, etc.).
• Antibody format (e.g. complete antibody, Fab, scFv, etc.) For complex antibodies, including bispecific or multispecific antibodies, be as specific as possible (e.g., scFv fusion to the beginning of the VH domain, IgG format with two different light chains and two different heavy chains with electrostatic mutations...).
• Name and/or structure of targeted antigen.
• Expression system.
• Clone name(s) and laboratory code name(s).
• Source of the original antibody (or antibodies) that provided the binding affinity [e.g., hybridoma (including species origin such as mouse, rat, etc.)].
• If appropriate, the closest human V, J and C genes and alleles (results obtained with IMGT/Domain Gap/Align tool).
• Subsequent engineering of V domains. e.g., none, humanization by CDR grafting (specify the CDR definition that was used), humanization by resurfacing framework mutations, etc.
• If the terminal lysine is absent in the heavy chain amino acid sequence, a statement confirming that there is no lysine codon in the precursor nucleotide sequence. If this is not the case, the lysine should be added in the amino acid sequence with a statement that it is clipped.
• A list of all engineered mutations in the constant region, their locations, and their purpose.
• A graphic representation/drawing of the arrangement of the domains or linkage of the domains.
• For Antibody-drug Conjugates
  • Editable ChemDraw file of the linker/payload combination showing which parts of the molecule are the linker and which are the payload.
    • Sites of attachment of the linker to the monoclonal antibody included in the MS-Word sequence document.
    • The average number of payload/linker molecules conjugated to the monoclonal antibody, or a range if it is variable.
• For pegylated proteins
  • The details of pegylation: the end group and the polymer chain with the average number of repeat units (to 2 significant figures).
  • The details of the linker (not the reagent used): where the linker is attached to the active moiety, and, ideally, if multiple sites are involved, in what proportion they are modified.

All DNA and RNA-based substances

• The full nucleotide sequence of the substance in the following format: 50 nucleotides per line, in blocks of 10, with numbering at the end of each line, as editable text that can be copied. The nucleotide sequence should be annotated to delineate the relevant parts of the sequence (e.g., coding regions, control regions).
• A table of features providing an overview of the relevant parts of the sequence (not required for short oligonucleotides). The table should contain the annoation, a description of the annotation, the position and, color code used in the sequence. Where a new vector is derived from an existing one, a sequence alignment and a table of comparison should be provided.
• A schematic map of the entire nucleic acid showing inserted/deleted gene(s) and relevant functional parts (not required for short oligonucleotides).
• For pegylated substances
  • Details of pegylation; the end group and polymer chain with the average number of repeat units (to 2 significant figures).
  • Details of the linker (not the reagent used): where the linker is attached to the active moiety, and, ideally, if multiple sites are involved, in what proportion they are modified.
• For conjugated nucleic acid-based substances, the mean numbers of the conjugated part, and if known, positions where the conjugate is attached.

Cell therapies

Also called cell-based therapies and cell-based gene therapy substances

• Laboratory code names and/or other code names used in publications and clinical trials.
• Cell Source or Tissue of Origin.
  • Where a cell therapy substance is prepared from a body fluid-derived source (e.g., peripheral blood, umbilical cord blood), a description of the starting cell material should be provided, for example, isolation from peripheral blood or apheresis material, with characterization of the relevant cellular subpopulations within the starting material. Generally, this would include details of all cellular subpopulations that could be of clinical significance (e.g., undifferentiated stem cells or lymphocytes).
  • Where a starting cell material is derived from a cell bank (e.g., an iPSC derived cell line), information on the origin of the cell bank and characterization should be provided (examples are remumiocel (126) or raguneprocel (126)).
  • Where a cell substance is initially extracted from tissue, for example adipose tissue or a solid tumor, a comprehensive description of the starting material (e.g. primary or metastatic tumor site or metastatic lymph node), should be provided, as well as the harvest modality (e.g. surgical specimen or biopsy) and the detailed process for extraction of the cells. Characterization of the relevant isolated cell populations should also be provided.
• Key steps of the Manufacturing Process, including any manipulations.
  • Describe any cell enrichment or purification/selection of the starting or intermediate material that is performed at any step in the preparation of the cell substance. The reagents used need to be described. Reference to a tradename or proprietary substance, solution, or process should be avoided.
    • A list and detail any in vitro culture conditions, including cell activation and differentiation, number of passages, and/or population doublings. This must include any reagents, growth factors, cytokines or other additives to the culture media, coatings on the culture vessels, or other critical factors used during the manufacturing process.
  • For cell-based gene therapy substances (obtained by gene addition and/or gene editing), provide details of the methods used for the genetic modification of the cells, including descriptions of how the modifications are achieved.
  • Any in-process holding steps and intermediate or finished substance storage conditions, if applicable.
• Characterization/description of the Cell Substance
  • A detailed description of the final cell substance and the release criteria should be provided. This includes the identity and purity of all cell populations relevant to the biological function and critical cell impurities and extends to the activation state of the critical cell type(s), whether they have been antigen loaded, and their biological activity. The identity of the cell populations responsible for the biological activity should be described at the phenotypic level (cell surface expression profile using accepted surface markers).
  • Describe the final cell substance at a functional level, defining the key biological activities such as biomolecule release and/or functional assays (e.g., potency assays), including detailed descriptions of the test methods.
  • Where the substance is claiming to be a stem cell to act therapeutically, additional in vitro and/or in vivo information must be provided to demonstrate that the cells are capable of self-renewal, are unspecialized, and the population can give rise to a number of specialized cell types.
  • Where the substance is claiming to be composed of stem or progenitor cells to act therapeutically, additional in vitro and/or in vivo data must be provided to demonstrate the claimed cell functionality/ies.
  • Where the substance is claiming to be composed of stromal cells to act therapeutically, additional in vitro and/or in vivo data must be provided to demonstrate the claimed cell functionality/ies (e.g., biomolecule release).
• Genetic Manipulation. A detailed description of the vector and its insert or any other nucleic acid involved in the manipulation should be provided.
  • The full nucleotide sequence of any nucleic acids used during the manipulation process should be provided in the following format: 50 nucleotides per line, in blocks of 10, with numbering at the end of each line, as editable text. The nucleotide sequence should be annotated to delineate relevant parts of the sequence.
  • A table of features providing an overview of the relevant parts of the sequence. The table should contain the annotation, a description of the annotation, the position, and the color code used in the sequence. Where a new vector is derived from an existing one, a sequence alignment and table of comparison should be provided.
  • Any impact on the characteristics of the cell population (e.g., through modification of a known gene function) should be described in line with the section on ‘characterization/description’.
  • Where applicable, provide information on pseudotyping of any vectors used.