Chromatography column qualification in manufacturing methods for producing anti-TNF antibody compositions

A method of operating a chromatography column in methods of manufacture for producing anti-TNF antibodies,

• wherein the anti-TNF antibodies comprise a heavy chain (HC) comprising amino acid sequence of SEQ ID NO:1 and a light chain (LC) comprising amino acid sequence of SEQ ID NO:2 or an antigen binding fragment thereof,
• said method comprising:
• collecting a column outlet signal and accumulated flow parameters at two or more intervals of at least one mobile phase transition front during a first operation of the chromatography column comprising column packing;
• determining a model gamma cumulative distribution curve based on the collected column outlet signal and accumulated flow parameters for the at least one mobile phase transition front using Formula Ia for a rising transition front or Formula Ib for a falling transition front,
• C
• =
• 1
• -
• 1
• Γ
• ⁡
• (
• k
• )
• ⁢
• γ
• ⁡
• (
• k
• ,
• V
• -
• V
• i
• θ
• )
• ⁢
• ⁢
• or
• Formula
• ⁢
• ⁢
• Ia
• C
• =
• 1
• Γ
• ⁡
• (
• k
• )
• ⁢
• γ
• ⁡
• (
• k
• ,
• V
• -
• V
• i
• θ
• )
• Formula
• ⁢
• ⁢
• Ib
• wherein C is the column outlet signal for a given V, V is the accumulated flow divided by column volume, and k, θ, and Vi are the shape, scale, and offset parameters, respectively, used to define the curve;
• calculating a height equivalent theoretical plate (HETP) value for the at least one mobile phase transition front using Formula II and the model gamma cumulative distribution curve parameters of k, θ, and VI,
• HETP
• =
• σ
• 2
• μ
• 2
• ⁢
• L
• Formula
• ⁢
• ⁢
• II
• wherein
• μ=kθ+Vi
• σ=√{square root over (kθ2)}
• L=column length
• assessing the quality of the chromatography column packing based on said calculated HETP value;
• wherein the quality of the chromatography column packing is acceptable if the calculated HETP value is above a lower control limit and below an upper control limit,
• wherein the lower control limit and the upper control limit are determined using a compiled trend of determined HETP values;
• or
• the method further comprises conditioning, replacing, or repacking the chromatography column to improve the quality of the chromatography column packing if the calculated HETP value is below the lower control limit or above the upper control limit.