I’m gonna need a physique check bro
Zijn geen Belgen die Daan heten
(Ignore this message if you don't care about in-depth HGH quality control)
I'm not a chemist, so I can't make HGH myself, but I know what the steps are and where you can go wrong.
Step 1: The human growth hormone gene is inserted into bacteria (E. coli). The bacteria take the gene in and start producing GH. However, this form of GH is not folded correctly yet. E. coli cannot do that. Instead, they produce inclusion bodies (insoluble protein clumps).
Step 2: The bacteria need to be broken open (it's called lysis). The inclusion bodies are then isolated. These clumps contain GH but in a denatured, inactive form.
Step 3: The inclusion bodies have to be dissolved by using strong denaturants (substances that disrupt the structure and function of proteins or nucleic acids without breaking their primary chemical bonds). Guanide hydrochloride, urea and detergents are used for this. When done correctly, the GH gets unfolded. Imagine your ear wires in your pockets that are all tangled up, they get straightened into a long straight line.
Step 4: Now the GH needs to be refolded to the correct 3D structure, I.e. its final form. This is where it often goes wrong. Why? Because it needs the exact temperature, precise redox environment, controlled pH, correct buffer strength, correct disulfide bond pairing, slow and controlled dilution and chaperone assisted folding (those are proteins that help make the GH fold correctly).
A lot can go wrong here. This process can be done too quickly, low-cost buffers can be used, absence or too few chaperone proteins to make sure the GH get folded correctly are present, there can be uncontrolled oxygen exposure, no controlled disulfide bond pairing (HGH consists of 2 parts, bound by a disulfide bond, if this bond gets 'damaged' then the HGH will be inactive). Another common mistake is refolding in bulk (kilograms at once). All these faults can lead to a large fraction of misfolded HGH, which is biologically inactive.
After the manufacturing of HGH, freeze-drying at incorrect temperatures causes denaturation (losing its correct 3D structure) of HGH. Having wrong excipients (inactive ingredients such as binding substances to keep the active substance together) can contaminate the product and cause denaturation. There needs to be proper quality control to make sure the wrong substances don't get in contact with the HGH. Moisture can denature HGH. Wrong pH or even pH shifts denatures HGH. Exposure to oxygen leads to oxidation (losing a hydrogen atom), denaturing the HGH.
These are common mistakes when manufacturing HGH.